Objective To explore the ultrasound imaging characteristics of thyroid papillary carcinoma complicated cervical lymph node tuberculosis, and to elucidate the key points of ultrasound diagnosis and to distinguish with cervical lymph node metastasis of papillary thyroid carcinoma.Methods In total, 1 5 well-documented cases of papillary thyroid carcinoma diagnosed definitely were selected, and there were 6 cases of concomitant lymph node metastasis. The ultrasonography of lymph node enlargement was analyzed, and the differences of the ultrasonographic characteristics between lymph node tuberculosis and metastatic lymph node including the location, swelling, calcification, blood flow and regional nodal liquefaction. Results Thyroid papillary carcinoma complicated with cervical lymph node tuberculosis was often found in the areas of Ⅲ,Ⅳ and Ⅴ, especially in the area of Ⅴ. Variety of echo was mixed in tuberculous of lymph node, and the echo was inhomogenous. The tuberculosis of lymph node calcification was patchy inhomogeneous distribution.The echo in part of liquefaction of lymph node tuberculosis was cottony weak. The flow signal of tuberculous lymph appeared the surrounding or internal punctate distribution,and the soft tissue was echogenic and disorder around the lymph node tuberculosis. Conclusion When ultrasonography examination is performed in the patients with the thyroid papillary carcinoma complicated with cervical lymph node enlargement, the history should be considered to analyze the ultrasound characteristics to dignose by observing the lesions of the surrounding soft tissues.

Objective To explore the effect of Numb on tubular epithelial-to-mesenchymal transition (EMT) in rat proximal epithelial cells. Methods NRK52E cells were treated with different concentrations of recombinant human transforming growth factor-β1 (TGF-β1) (0, 1, 5, 10, 15, 20 μg/L) for 48 h or 10 μg/L TGF-β1 for different times (0, 24, 48, 72 h) in vitro. The expressions of E-cadherin, a-smooth muscle actin(α-SMA) and Numb in NRK 52E cells were detected by RT-PCR, Western blot and immunofluorescence staining. Meanwhile Numb siRNA oligo was transfected into NRK 52E cells with lipofectamine before TGF-β1 treatment, then Western blot was applied to detect the protein expression of E-cadherin, α-SMA and Numb in NRK52E cells. Results TGF-β1 could induce EMT in NRK52E cells in dose- and time-dependent manner. During the progress of TGF-β1-induced EMT, the protein expression of Numb in 5, 10, 15, 20 μg/L group was 1.33 folds (P=0.024), 1.39 folds (P=0.035), 1.45 folds (P=0.025), 1.51 folds (P=0.000) respectively as compared to 0 μg/L group. Likewise, the protein and mRNA expression of Numb in 24 h, 48 h, 72 h group was 1.48 folds (P=0.046) and 1.56 folds (P=0.012), 1.54 folds (P=0.011) and 1.82 folds (P=0.008), 1.79 folds (P=0.028) and 1.82 folds (P=0.002) respectively as compared to 0 h group. Moreover, large amount of Numb was accumulated in the cytoplasm. Down-regulation of Numb expression by siRNA transfection did not influence the basal expression of E-cadherin and α-SMA in NRK 52E cells, but attenuated the progression of EMT in NRK52E cells induced by TGF-β1. The up-regulation of α-SMA protein was reduced to 18.1% (P=0.004) while the down-regulation of E-cadherin protein was reversed to 2.19 folds (P=0.004). Conclusion Numb can promote EMT in rat proximal epithelial cells.

AIM: To observe the expression of neuronal Aryl hydrocarbon receptor nuclear translocator 2 (ARNT2) involved in neuronal apoptosis evoked by low K + and to investigate the relationship between ARNT2 and neuronal apoptosis. METHODS: After neuron apoptosis model was established, the changes of mRNA and protein of ARNT2 during apoptosis were investigated by RT-PCR and Western blotting, respectively. Immunofluorescence was analyzed by confocal microscopy to probe the subcellular localization of ARNT2. RESULTS: Induced by low K +, the expression of ARNT2 mRNA was up-regulated obviously at the point of 30 min, and peaks at the point of 1 h. This up-regulated expression lasted for 12 h, and the variation of ARNT2 protein was similar to that of mRNA. The results of immunofluorescence analyzed by confocal microscopy showed that the localization of ARNT2 protein was in the nucleus. CONCLUSION: ARNT2 locate in nuclei of normal cerebellar granule neurons of rat. During the process of apoptosis evoked by low K +, both mRNA and protein of ARNT2 are overexpressed.

AIM: To analyze the subcellular localization of Arnt2 in rat cerebellar granule neurons (CGNs). METHODS: Based on the amino acids sequence of Arnt2 (LOCUS:NP_036913), the subcellular localization of Arnt2 in eukaryotic cells and the nuclear export signals (NES) of Arnt2 were predicted in CBS bioinformatics database. The subcellular localization of Arnt2 in rat cerebellar granule neurons was detected by the method of laser scanning confocal microscopy (LSM) analysis. RESULTS: It was predicted that Arnt2 located in nuclei of eukaryotic cells with the most probability, while located in cytoplasmic mitochondria with a slight possibility. A nuclear export signal was found in Arnt2 amino acids sequence, it was identified to be the leucine of No.143 that located in N-terminal of Arnt2 amino acids sequence. Finally, the result of LSM analysis shows nuclear localization of Arnt2 in rat CGNs. CONCLUSION: Arnt2 is located in nuclei of normal rat CGNs, it suggests that Arnt2 has the tendency to translocate into mitochondria after induced by some of inducible factors, for both the possibility of mitochondria localization and NES exist in Arnt2 amino acids sequence.

Objective To investigate the pathological type and characteristics of renal allograft in kidney transplantation recipients,and to analyze the relevant clinical conditions and prognosis of renal function.Methods 230 patients received renal allograft biopsy after renal transplantation.The pathological type and characteristics of renal allograft specimens were observed,and the serum creatinine (SCr) in the recipients with different pathological types were analyzed.The function of renal allograft in the recipients was followed-up after one year,and their prognosis was evaluated.Results In 10 cases of protocol biopsy,normal renal tissues were found in 9 cases,IgA nephropathy occurred at the 3rd month after transplantation.In 220 cases having impaired renal function,there were 33 cases of borderline change,45 cases of acute rejection (AR),24 cases of chronic rejection (CR),26 cases of chronic allograft nephrapathy (CAN),and 39 cases of posttransplantation glomerulonephritis (PTGN).Except for above 167 cases,lesions of 28 cases showed multiple pathology types.Furthermore,there were 8 cases of calcineurin inhibitor nephrotoxicity (CNI-NT),7 cases of BK virus nephropathy (BKVN),and 5 cases of acute tubular necrosis (ATN).Five cases could not be diagnosed for little tissue.In the recipients with pathological diagnosis of borderline change,AR,CR,CAN and nephritis,SCr levels were (171 ± 17),(259 ± 25),(343 ± 33),(406 ± 67) and (207 ± 26) respectively.There was significant difference in SCr levels of recipients among the above 5 groups (P＜0.01).One year after biopsy,137 recipients (80.2%) were followed up.The dysfunction rate of renal allograft was 3.1%,18.2%,22.2 %,33.3% and 13.5% respectively.The △SCr was (-47 ± 20.7),(-37.3± 36.9),(25.5 ± 24.3),(13.5 ± 27.7) and (25.2 ± 17.1) μmol/L respectively.Conclusion Complex and diverse pathological changes were showed in renal allograft.Accurate diagnoses come from renal biopsy and clinical analysis may help clinicians select appropriate treatment programs to promote long-term graft survival.

Objective:To analyze and summarize the characteristics of ultrasonographic images of non-lactating mastitis, improve the diagnostic accuracy, and guide the clinical treatment.Methods:The ultrasonic images of 46 patients with non lactation mastitis diagnosed by pathology in People′s Hospital of Heyuan from January 2017 to November 2020 were retrospectively analyzed. The ultrasonic images were compared with the pathological results, and the ultrasonic characteristics of non lactation mastitis were analyzed.Results:Among the 46 cases of non lactation mastitis, there were 24 cases of chronic mastitis, 15 cases of granulomatous lobular mastitis, 5 cases of plasma cell mastitis and 2 cases of lymphocytic lobulitis. The ultrasonographic features of non lactation mastitis were patchy hypoechoic mixed with anechoic areas with poor sound transmission. Granulomatous lobular mastitis was easy to be misdiagnosed as malignant lesions because of its disordered echo.Conclusions:Mastering the ultrasound characteristics of non-lactation mastitis can help to improve the accuracy of diagnosis of non-lactation mastitis.

Objective To investigate the effects of fibrin-derived peptide Bβ15-42(FgBβ15-42)on mitophagy in acute kidney injury(AKI)with ischemia-reperfusion at the cellular level.Methods Human kidney-2(HK2)cells were cultured in vitro and ran-domly divided into five groups:normal control group(Control group),hypoxia/reoxygenation(H/R)model group(H/R group),H/R+9μmol/L FgBβ15-42 peptide group(Fg group),H/R+20μmol/L chloroquine group(CQ group),and H/R+9μmol/L FgBβ15-42 peptide+20μmol/L chloroquine group(Fg+CQ group).Control group:cells were homogenized and replaced with complete medium and incubated in an aerobic incubator(5％CO2,21％O2)for 28hours.H/R group:cells were added to sugar-free and serum-free DMEM/F12medium and then placed in a hypoxic incubator(5％CO2,1％O2 and 94％N2)for 24hours,washed with PBS and added to DMEM/F12medium containing serum and then placed in a reoxygenated environment(5％CO2,21％O2)for 4hours.Fg group:mod-eled according to the above method,intervention was performed by adding FgBβ15-42 peptide 9μmol/L to the complete medium when reoxygenation was performed after the completion of hypoxia.CQ group:pre-treatment with chloroquine(20μmol/L)in serum-free medium at the time of homogenization,with the same post-procedural steps as in the H/R group.Fg+CQ group:the intervention was performed according to the dosing sequence of the CQ group and FgBβ15-42 group at different time periods as mentioned above,and the modeling time of the H/R group.After the intervention,CCK-8 was used to detect the cell viability of HK2.The mitochondrial morphol-ogy,mitophagosome structure and number were observed by electron microscope.The expression levels of autophagy related proteins p62 and LC3 were detected by Western blot.Results Compared with the control group,the survival rate of H/R group was decreased(P＜0.01);the mitochondrial damage was obvious,such as swelling and spinous rupture,and the number of mitophagosomes increased.The expression of autophagy protein LC-3 Ⅱ/Ⅰ was up-regulated(P＜0.05),and the expression of autophagy substrate protein p62 was down-regulated(P＜0.01).Compared with H/R group,the survival rate of Fg,CQ and Fg+CQ groups increased(P＜0.01),mito-phagosome was decreased.the expression of LC-3 Ⅱ/Ⅰ was down-regulated(P＜0.01),the expression of p62 was up-regulated(P＜0.05),and autophagy was inhibited.Conclusion The intervention of FgBβ15-42 peptide can reduce the level of autophagy and mitophagy in H/R-AKI cells,and has a protective effect on H/R injury in HK2 cells.

Objective:To report a rare case of paroxysmal nocturnal hemoglobinuria (PNH) complicated with chronic tubulointerstitial nephropathy, combined with literature review, and discuss the clinical, imaging and pathological characteristics of the disease and the diagnosis and treatment ideas.Methods:The patient's clinical data, magnetic resonance imaging (MRI) and kidney pathological examination results, treatment measures and effects were collected and reported. Through systematic review of relevant literature, the clinical manifestations and pathogenesis of chronic tubular interstitial nephropathy complicated by PNH were summarized and discussed.Results:In this case, PNH was diagnosed for more than 30 years, the peripheral blood PNH clone was positive, urine specific gravity was 1.012, urine pH 6.0-7.0, urine protein (+), urine sugar (3+), serum creatinine 259 μmol/L, serum lactic acid dehydrogenase 800 U/L. MRI showed bilateral renal cortical signal was low intensity on both T1- and T2- weighted images. Kidney biopsy revealed remarkable chronic tubulointerstitial nephropathy with massive hemosiderin deposition in proximal tubular cells demonstrated by Prussian blue staining and electron microscopy. By using low-dose prednisone to control hemolytic attack and other supportive treatments, the patient's renal function has been stabilized for a long time.Conclusions:PNH complicated with chronic tubulointerstitial nephritis is easy to be misdiagnosed due to insidious onset. MRI and kidney histopathological examination are helpful to clarify the diagnosis. Early diagnosis and treatment are helpful to improve the prognosis of such patients.

BACKGROUND:Several studies have found that the total flavonoids of rhizome drynariae can promote neovascularization in the induced membrane,improve the biological properties of the induced membrane,and accelerate bone remodeling in the induced membrane,but the related molecular mechanisms still need to be further explored. OBJECTIVE:To explore the effect of total flavonoids of rhizome drynariae on bone remodeling in rat femoral Masquelet-induced membrane model by regulating H-type blood vessels. METHODS:Thirty-six male Sprague-Dawley rats were stratified by body mass and then randomly divided into blank group,model group and traditional Chinese medicine group,with 12 rats in each group.A 4-mm femoral bone defect model was established in all the rats.Bone defects in the model group and traditional Chinese medicine group were filled with polymethylmethacrylate bone cement.At 6 weeks after modeling,the tail bone of the rats was implanted in the blank group,as well as in the other two groups after removal of bone cement.The traditional Chinese medicine group was given 157.5 mg/kg per day of total flavonoids of rhizome drynariae at 3 days after bone implantation,while the model and blank groups were given the same amount of saline by gavage until the 8th week after bone implantation.Bone graft samples were taken for relevant testing at 8 weeks after implantation. RESULTS AND CONCLUSION:X-ray films showed that in the blank group,the fracture line in the defect area was clear,and only a small amount of bone callus formed;in the model group,the bone defect area still existed,where discontinuous cortical bone was visible;in the traditional Chinese medicine group,the defect area was filled with newborn bone tissues,the bone marrow cavity and part of the cortical bone formed,and the fracture line disappeared.Micro-CT scans showed that the amount of new bone in the defect area was low in the blank group,the number of bone trabeculae in the defect area was significantly increased in the model group,and a large amount of new bone tissue was filled in the bone defect area in the traditional Chinese medicine group.Hematoxylin-eosin staining results showed that in the blank group,only a small amount of new bone formed in the defect area and the quality of osteogenesis was poor;in the model group,there was more new bone tissue in the defect area,but some fibrous connective tissues were interspersed within the bone tissue;and in the traditional Chinese medicine group,a large amount of new bone formed in the defect area and the quality of osteogenesis was the best.CD31/Emcn immunofluorescence double-labeling staining results showed that the number of H-type blood vessels in the newborn bone tissue in the bone defect area of the blank group was sparse and sparsely distributed;compared with the blank group,there were more H-type blood vessels in the bone tissue in the bone defect area of the model group,and the blood vessels were distributed in relatively regular strips;the number of H-type blood vessels in the bone defect area of the traditional Chinese medicine group was the highest and the blood vessels were densely distributed.To conclude,the total flavonoids of rhizoma drynariae can upregulate the expression of H-type blood vessels to enhance the angiogenic-osteogenic effect,improve the osteogenic efficiency of the rat femoral Masquelet induced membrane model,and promote bone remodeling.

OBJECTIVE: To develop a drug-loaded composite microsphere that can simultaneously release the berberine (BBR) and naringin (NG) to repair infectious bone defects. METHODS: The NG was loaded on mesoporous microspheres (MBG) to obtain the drug-loaded microspheres (NG-MBG). Then the dual drug-loaded compound microspheres (NG-MBG@PDA-BBR) were obtained by wrapping NG-MBG with polydopamine (PDA) and modifying the coated PDA with BBR. The composite microspheres were characterized by scanning electron microscopy, X-ray diffraction, specific surface area and pore volume analyzer, and Fourier transform infrared spectroscopy; the drug loading rate and release of NG and BBR were measured; the colony number was counted and the bacterial inhibition rate was calculated after co-culture with Staphylococcus aureus and Escherichia coli for 12 hours to observe the antibacterial effect; the biocompatibility was evaluated by live/dead cell fluorescence staining and cell counting kit 8 assay after co-culture with rat's BMSCs for 24 and 72 hours, respectively, and the osteogenic property was evaluated by alkaline phosphatase (ALP) staining and alizarin red staining after 7 and 14 days, respectively. RESULTS: NG-MBG@PDA-BBR and three control microspheres (MBG, MBG@PDA, and NG-MBG@PDA) were successfully constructed. Scanning electron microscopy showed that NG-MBG@PDA-BBR had a rough lamellar structure, while MBG had a smooth surface, and MBG@PDA and NG-MBG@PDA had a wrapped agglomeration structure. Specific surface area analysis showed that MBG had a mesoporous structure and had drug-loading potential. Low angle X-ray diffraction showed that NG was successfully loaded on MBG. The X-ray diffraction pattern contrast showed that all groups of microspheres were amorphous. Fourier transform infrared spectroscopy showed that NG and BBR peaks existed in NG-MBG@PDA-BBR. NG-MBG@PDA-BBR had good sustained drug release ability, and NG and BBR had early burst release and late sustained release. NG-MBG@PDA-BBR could inhibit the growth of Staphylococcus aureus and Escherichia coli, and the antibacterial ability was significantly higher than that of MBG, MBG@PDA, and NG-MBG@PDA ( P<0.05). But there was a significant difference in biocompatibility at 72 hours among microspheres ( P<0.05). ALP and alizarin red staining showed that the ALP positive area and the number of calcium nodules in NG-MBG@PDA-BBR were significantly higher than those of MBG and NG-MBG ( P<0.05), and there was no significant difference between NG-MBG@PDA and NG-MBG@PDA ( P>0.05). CONCLUSION NG-MBG@PDA-BBR have sustained release effects on NG and BBR, indicating that it has ideal dual performance of osteogenesis and antibacterial property.
Rats ; Animals ; Osteogenesis ; Delayed-Action Preparations/pharmacology* ; Microspheres ; Berberine/pharmacology* ; Anti-Bacterial Agents/pharmacology* ; Escherichia coli