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Objective To investigate the role of fibroblast growth factor-23 (FGF23) in secondary hyperparathyroidism (SHPT). Methods (1)Serum FGF23 and intact parathyroid hormone (iPTH)from 38 maintenance hemodialysis (MHD) patients were measured by ELISA and chemiluminescence enzyme immunoassay respectively.(2) Parathyroid cells from six SHPT patients underwent parathyroidectomy with forearm autotrlansplantation were cultured for 24 h,then were induced by 0.1 mg/L FGF23.The supernatant was collected at 0.6,12,24 and 48 h respectively. The concentration of iPTH was measured by chemiluminescence enzyme immunoassay. (3)Protein expression of Klotho,FGFR1,FGFR3,GATA-3 and PCNA in parathyroid tissue from 33 SHPT eases and 3 healthy people were detected by immunohistochemistry SP and PV methods respectively. Positive cell rate and absorbance were calculated. Results (1) Serum FGF23 [(3901.85±2618.11) ng/L] was positively correlated with serum iPTH [(460.00±489.77) ng/L] in MHD patients. (2) 0.1 mg/L FGF23 suppressed iPTH secretion of parathyroid cells only at 24 h time point in vitro (P<0.05). (3) Expression of GATA-3, FGFR3, Klotho and PCNA was significantly increased and FGFRl was signiticantly decreased in parathyroid tissue of SHPT-patients as compared to healthy people. (4) Positive cell rate of GATA-3 was positively correlated with iPTH (r~2=0.1901, P=0.0425) and PCNA (r~2=0.2584, P=0.0025). Klotho was positively correlated with FGFRI and FGFR3 (r~2=0.2046, P=0.0082;r~2=0.2833, P=0.0014). PCNA was negatively correlated with FGFR1 (r~2=0.1292, P=0.0399) and positively correlated with FGFR3 (r~2=0.1226, P=0.0457). FGFR1 was negatively correlated with serum phosphate (r~2=0.2329, P=0.0044) and positively correlated with serum calcium (r~2=0.1422, P=0.0305). Conclusions FGF23 level is positively correlated with iPTH level in MHD patients. FGF23 can inhibit iPTH secretion of parathyroid cells in a weak and short way, which may be associated with the proliferation of GATA-3 positive cells and parathyroid cells, the up-regulation of FGFR3 and the down-regulation of FGFR1 expression.

Objective To observe the steps of vascular smooth muscle cells (VSMCs) calcification induced by high phosphate enviroment in vitro. Methods VSMCs were incubated with high phosphate (2.5 mmol/L or 3.5 mmool/L) medium for different times. Expression of core binding factor α1(Cbfα1), osteopontin(OP), collagen type Ⅰ(Col Ⅰ), osteocalcin(OC) and α-smooth muscle actin (α-SMA) was investigated by Western blot, immunofluorcscencc staining and real time PCR. Mineral deposition was assessed by von Kossa aad Alizarin red staining. Ultrastructure of VSMCs calcification was observed by electron microscopy (EM). Results Up-regulated expression of osteoblast-specific transcription factor Cbfα1 in the nuclei oceured at as early as 12 hours. The protein of Col Ⅰ and OP was up-regalated when VSMCs were incubated in high phosphate medium for 3 days, and content of OC increased at the time of 6 days. When cultured in 2.5 mmol/L phosphate medium for 15 days, VSMCs lost their lineage marker α-SMA, developed granular calcium deposits. Moreover, the results of real time PCR indicated mRNA level of OP and Col Ⅰ increased at day 1, OC increased at day 5 and α-SMA level decreased at day 10, respectively. Ultrastructural analysis also confirmed the presence of collagen and matrix vesicles in the cells. Conclusion VSMCs phenotype transformation induced by high phosphate enviroment is an orchestrated, highly regulated process.

Objective To investigate the significant of peripheral CD4+CD69+T lymphocytes in patients with autoimmune hemolytic anemia (AIHA)/Evans syndrome (ES).Methods In this study peripheral blood samples from 32 patients with AIHA/ES (15 hemolytic episode patients,17 remission patients) and 13 healthy controls were collected.Patients with AIHA/ES were recruited in Tianjin Medical University General Hospital from October 2015 to May 2016.The percentages of CD69+ T lymphocytes were analyzed by flow cytometry.The expression of CD69 mRNA in CD4+T lymphocytes which was sorted from peripheral blood by magnetic activated cell sorting (MACS) was detected using real-time PCR.Soluable CD69 was measured by ELISA.Results In hemolytic episode patients,the ratio of CD3+CD69+/CD3+T lymphocytes [(3.08 ± 1.48)％] was significantly higher than that in healthy controls [(1.28 ± 0.83)％,P＜0.01] and in remission group[(1.96± 1.33)％,P＜0.05].The absolute count of CD3+CD69+T lymphocytes in hemolytic episode group [(2.94± 1.81)× 107/L] was higher than that in healthy controls [(1.48± 1.42)× 107/L,P＜0.05].The ratio of CD3+CD4+CD69+/CD3+CD4+T cells in hemolytic episode group [(2.16± 1.56)％] was significantly higher than that in remission group [(1.16±0.62)％,P＜0.05] and healthy controls[(0.94±0.78)％,P＜0.05].The quantity of CD3+CD4+CD69+T lymphocytes in hemolytic episode group[(1.04±0.98)× 107/L] was higher than in healthy controls [(0.44± 0.38) × 107 / L,P＜0.05].The ratio of CD3+CD8+CD69+/CD3+ CD8+T lymphocyte in hemolytic episode group [(4.87±2.56)％] was significantly higher than that in healthy controls[(1.83± 1.27)％,P＜0.01].The quantity of CD3+CDs+CD69+T lymphocytes in three groups did not show significant difference.The ratio of CD3+CD4+CD69+/CD3+CD4+T lymphocytes in hemolytic episode group was negatively correlated with hemoglobin (Hb) (P＜0.01),positively correlated with the percentage of reticulocytes (Ret％)(P=0.01)total bilirubin(TBil),indirect bilirubin(IBil) (P＜0.01) and not correlated with absolute reticulocytes count,lactic dehydrogenase (LDH),complement 3(C3),complement 4 (C4).The ratio of CD3+CD4+CD69+/CD3+CD4+T lymphocytes in remission group was negatively correlated with Hb (P＜0.05).In hemolytic episode patients CD69 mRNA (32.26±35.11) was significantly higher than that in remission group(6.05±5.87)(P＜0.05)and healthy controls (1.76± 1.85)(P＜0.01).CD69 mRNA in remission group was significantly higher than healthy controls (P＜0.05).Serum CD69 in hemolytic episode patients [(494.21 ± 16.06) ng/L] was significantly higher than that in healthy controls [(441.39± 104.6) ng/L,P＜0.05].Conclusion Our findings suggest that the proportion of CD4+CD69+ T lymphocytes increase in AIHA/ES patients,which is correlated with the severity of disease.

Enrichment of trace bioactive constituents and metabolites from complex biological samples is chal-lenging. This study presented a one-pot synthesis of magnetic polydopamine nanoparticles (Fe3O4@-SiO2@PDA NPs) with multiple recognition sites for the magnetic dispersive solid-phase extraction (MDSPE) of ginsenosides from rat plasma treated with white ginseng. The extracted ginsenosides were characterized by combining an ultra-high-performance liquid chromatography coupled to a high-resolution mass spectrometry with supplemental UNIFI libraries. Response surface methodology was statistically used to optimize the extraction procedure of the ginsenosides. The reusability of Fe3O4@-SiO2@PDA NPs was also examined and the results showed that the recovery rate exceeded 80％ after recycling 6 times. Furthermore, the proposed method showed greater enrichment efficiency and could rapidly determine and characterize 23 ginsenoside prototypes and metabolites from plasma. In com-parison, conventional methanol method can only detect 8 ginsenosides from the same plasma samples. The proposed approach can provide methodological reference for the trace determination and charac-terization of different bioactive ingredients and metabolites of traditional Chinese medicines and food.

Carcinoid is a rare clinical disease, especially primary carcinoid of testis. On December 11, 2019, a patient with primary testicular carcinoid was admitted to the Fourth Hospital of Hebei Medical University. The physical examination, color Doppler ultrasound and scrotal CT showed that left testicular tumor. The patient underwent radical orchiectomy and sent for pathology examination on December 14, 2019. Gastroscopy, enteroscopy and CT examination of pelvic and abdominal cavity showed no obvious abnormalities. Postoperative pathology combined with immunohistochemistry was diagnosed as primary carcinoid of testis. The patient was followed up for 23 months with no recurrence.

Objective To evaluate the additional role of 99mTc-MIBI SPECT-CT imaging before parathyroidectomy (PTX) for secondary hyperparathyroidism (SHPT) patients.Methods A total of 359 SHPT patients from the First Affiliated Hospital of Nanjing Medical University were enrolled in the study from April 2011 to January 2016.Patients were divided into two groups according to the difference of 99mTc-MIBI imaging techniques preoperatively.Patients in group A had dual-phase 99mTc-sestamibi parathyroid scintigraphy performed only.Patients in group B had SPECT-CT parathyroid scintigraphy added in the early phase.With the parathyroid glands confirmed by pathology after operation being the referencc,scnsitivity,specificity,consistency and surgical success rates of the patients in two groups were compared.Results 359 patients (166 men and 193 women) were enrolled,among which 339 on hemodialysis and 20 on peritoneal dialysis.The average age of those patients was (45.90± 11.20) years and median dialysis age was 84 (60.0,120.0) months.Total detection sensitivity,specificity,consistency in group A (169 patients) were 65.54％,56.52％,65.24％ and in group B (190 patients) were 73.84％,84.62％,74.21％ respectively.Total sensitivity and consistency were higher in group B than those in group A (P ＜ 0.01),while no difference in total specificity was observed between two groups (P=0.055).In group A,detection sensitivity of parathyroid glands were 47.56％,77.44％,57.14％ and 79.88％ in right upper gland (RU),right lower gland (RL),left upper gland (LU) and left lower gland (LL) respectively.Consistency in group A were 48.52％,77.51％,56.21％ and 78.70％ respectively.In group B,sensitivity were 58.33％,83.78％,69.78％ and 82.89％ in RU,RL,LU and LL respectively.Consistency in group B were 59.47％,84.21％,70.53％ and 82.63％ respectively.Sensitivity and consistency of upper glands were lower than lower glands in both groups (P ＜ 0.01).When SPECT-CT was added,sensitivity and consistency of upper glands for both sides were higher in group B than those in group A,while no difference of surgical success rates was found between two groups (87.57％ vs 92.63％,P=0.107).Conclusion 99mTc-MIBI SPECT-CT can be combined with anatomic image effectively and increase the sensitivity and consistency of total and upper parathyroid glands.It can also make the accurate location of the lesions,which improves the efficiency of the operation.

Objective:To observe heart rate circadian rhythm in patients with chronic kidney disease (CKD) stage 5 and to analyze the effects of parathyroidectomy (PTX) on heart rate circadian rhythm in severe secondary hyperparathyroidism (SHPT) patients.Methods:A cross-sectional observation was performed in 213 patients with CKD stage 5 and 96 controls, and the patients were divided into those with severe SHPT (PTX group, n=70) and without severe SHPT (non-PTX group, n=143). Forty-six PTX patients were followed up prospectively. The baseline data were compared among these groups. Holter electrocardiogram was performed for each participant. Non-dipping heart rate was defined as night/day heart rate ratio greater than 0.9. Multiple linear regression analysis was used to analyze the related factors of heart rate circadian rhythm in patients with CKD stage 5. Results:The 24-hour, daytime and nighttime mean heart rate in patients with CKD stage 5 were all higher than those in controls, especially in PTX group (all P<0.05). The night/day heart rate ratios of controls and CKD stage 5 patients were (0.81±0.08) and (0.91±0.08) respectively ( P<0.01). Correlation analysis showed 24-hour and daytime or nighttime mean heart rate in patients with CKD stage 5 were positively correlated with serum levels of phosphorus and ln(alkaline phosphatase), while nighttime mean heart rate and night/day heart rate ratio were positively related with serum intact parathyroid hormone level. After adjusting with postoperative follow-up period (median time: 10.9 months), 24-hour and nighttime mean heart rate, and night/day heart rate ratio in PTX patients all decreased significantly (all P<0.01). Conclusions:Heart rate is increased and circadian rhythm is abnormal in patients with CKD stage 5, which are related with mineral and bone disorder. PTX significantly decreases 24-hour and nighttime mean heart rate in severe SHPT patients, and improves the heart rate circadian rhythm.

Objective To investigate the influencing factors of hungry bone syndrome (HBS) in maintenance hemodialysis patients with secondary hyperparathyroidism (SHPT) after parathyroidectomy (PTX). Methods A retrospective study was conducted on maintenance hemodialysis patients with SHPT undergoing successful parathyroidectomy with autotransplantation. Clinical data and perioperative indicators of the selected patients were collected. The enrolled patients were divided into HBS group and non-HBS group based on whether the lowest level of blood calcium less than 2.0 mmol/L after surgery. The difference of general clinical data and perioperative indicators between the two groups were compared. The risk factors of HBS were analyzed by logistic regression analysis. Multiple linear regression method was used to analyze the independent factors affecting the maintenance time of intravenous calcium supplementation, the total amount of calcium supplementation during intravenous calcium supplementation and the highest serum level of potassium within 24 h after surgery. Results A total of 306 patients were included in the study. All patients had low levels of serum calcium after operation. There were 230 patients (75.16% ) with the lowest blood calcium＜2.00 mmol/L after PTX (HBS group), and 76(24.84%) cases in the non-HBS group. Predialysis coefficient of serum calcium=(preoperative blood calcium-2.20) mmol/L÷0.01 mmol/L. Logistic regression analysis showed that higher predialysis coefficient of serum calcium (B=-0.063, OR=0.939, 95% CI 0.894-0.987, P=0.013) and lower level of preoperative serum alkaline phosphatase (ALP) (B=0.035, OR=1.033, 95%CI 1.019-1.050, P＜0.001) were independent risk factors for HBS. Multiple linear regression analysis revealed that preoperative blood intact parathyroid hormone (iPTH) (B=0.017, P＜0.001 and B=0.041, P＜0.001), preoperative serum ALP (B=0.052, P＜0.001 and B=0.107, P＜0.001) and preoperative hemoglobin (Hb) (B=-0.453, P=0.041 and B=-1.058, P=0.007) were independent factors affecting the maintenance time of intravenous calcium supplementation and the total amount of calcium supplementation in HBS patients. Preoperative predialysis level of serum potassium (B=0.859, P＜0.001) was the independent influencing factor of the maximum level of serum potassium within 24 hours after surgery. Conclusions Patients with lower levels of preoperative serum calcium and higher levels of serum ALP are prone to HBS after PTX. Postoperative calcium supplementation may need to be strengthened in HBS patients with higher preoperative iPTH and ALP levels and lower preoperative Hb levels. High preoperative basal potassium levels may increase the risk of hyperkalemia after PTX.

Objective:To determine the prognostic values of clinical and laboratory features at the time of presentation on renal survival of patients with myeloperoxidase (MPO)-antineutrophil cytoplasmic antibody(ANCA)-associated glomerulonephritis (MPO-ANCA-GN).Methods:A total of 172 patients with MPO-ANCA-GN and hospitalized at the First Affiliated Hospital of Nanjing Medical University from January 2005 to December 2018 were enrolled. The baseline clinical characteristics and renal biopsy pathological data were analyzed, and the renal prognosis was followed up. The clinical and pathological characteristics of different renal prognosis in all patients and 112 patients who underwent renal biopsy were analyzed, and the related factors affecting renal survival were further discussed.Results:Among these 172 patients, 81 were males and 91 were females. The median serum creatinine at diagnosis was 343.7(174.2, 606.6) μmol/L and the median estimated glomerular filtration rate (eGFR) was 15.81(7.61, 38.04) ml·min -1·(1.73 m 2) -1. In total, 76 patients (44.2%) received initial renal replacement therapy (RRT). During a median follow-up duration of 20(3, 60) months, 73 patients (42.4%) progressed to end-stage renal disease (ESRD) and required dialysis, including 6 (8.2%) patients who entered RRT during follow-up and 67 (91.8%) patients who received RRT at the beginning. Among the 112 patients who underwent renal biopsy, the proportion of patients who progressed to ESRD in the sclerotic group was the highest (15/25, 60.0%). The baseline serum creatinine level ( P<0.001), urine red blood cell count ( P=0.012) and the proportion of glomerular sclerosis ( P=0.002) in the non-dialysis dependent group were significantly lower than those in the dialysis dependent group, while the levels of eGFR ( P<0.001), serum albumin ( P=0.002) and hemoglobin ( P<0.001) were higher than those of the dialysis-dependent group. Kaplan-Meier survival analysis showed that the renal survival rate of the focal group was the highest ( χ2=19.488, P<0.001, log-rank test), while the renal survival rate of the sclerotic group was significantly lower than that of the crescentic group ( χ2=5.655, P=0.017); higher levels of serum creatinine (>320 μmol/L, χ2=77.229, P<0.001) and urine red blood cell count (>300 cells/μl, χ2=8.511, P=0.004), lower levels of rheumatoid factor (<20 IU/ml, χ2=8.610, P=0.003), serum albumin (<30 g/L, χ2=11.060, P=0.001) and hemoglobin (<90 g/L, χ2=21.921, P<0.001) were associated with lower renal survival rate; in terms of treatment, the renal survival rate of the glucocorticoids plus mycophenolate mofetil group was significantly higher than that of the glucocorticoids plus cyclophosphamide ( χ2=5.056, P=0.025) or the glucocorticoids alone group ( χ2=16.459, P<0.001). Multivariate Cox regression showed that baseline serum creatinine >320 μmol/L ( HR=8.803, 95% CI 3.087-25.106, P<0.001) and serum albumin <30 g/L ( HR=2.566, 95% CI 1.246-5.281, P=0.011) were the related factors affecting renal survival. Conclusion:Serum creatinine and albumin levels of MPO-ANCA-GN patients at diagnosis may be the related factors that affect the patient's renal prognosis.