BACKGROUND: The osteoinduction by biomaterials has been proven in various animal experiments. OBJECTIVE: To investigate the osteoinduction of porous calcium phosphate cement on bone marrow mesenchymal stem cel s. METHODS: Bone marrow mesenchymal stem cel s were isolated from rabbits aged 1 week in vitro and labeled by PKH-67 or PKH-26, respectively. Forty-eight adult New Zealand white rabbits were randomized into four groups and porous calcium phosphate cement was implanted into both sides of gluteus maximus in each rabbit. Then, 1 mL PKH-26-labeled bone barrow mesenchymal stem cel s (1×1010/L) were injected into the superior gluteal artery branch at each side of gluteus maximus near the femur, and 1 mL PKH-67-labeled bone barrow mesenchymal stem cel s (1×1010/L) injected into tissues around the cement (group A); 1 mL PKH-26-labeled bone barrow mesenchymal stem cel s (1×1010/L) were injected into the each side of superior gluteal artery branch (group B); 1 mL PKH-67-labeled bone barrow mesenchymal stem cel s (1×1010/L) were injected into tissues around the cement (group C); the same amount of normal saline was injected into tissues around the cement (group D). At 3, 7 and 12 weeks after implantation, the cement and its surrounding tissues were extracted and detected by fluorescence microscope and Massion staining. Expression of bone morphogenetic protein 2 was analyzed by RT-PCR. RESULTS AND CONCLUSION: Under fluorescence microscope, PKH-26-labeled bone barrow mesenchymal stem cel s attached fast and distributed homogeneously; however, PKH-67-labeled bone barrow mesenchymal stem cel s attached slowly and exhibited a gradual y homogeneous distribution. Massion staining showed that ectopic new bone formation appeared to have an upward trend in al groups, and the area of new bones in groups A, B and C were larger than that of group D at different time points after implantation. There was a significantly higher expression of bone morphogenetic protein 2 in groups A, B and C compared with the group D at different time points after implantation (P < 0.05), and the expression was the highest in the group A (P < 0.05). In conclusion, the porous calcium phosphate cement can induce bone barrow mesenchymal stem cel s chemotaxis and osteogenetic differentiation. Besides, osteoblasts are differentiated from both the surrounding capil aries and body fluid, and capil ary-derived mesenchymal stem cel s occupy the important position.

Objective To investigate the significance of sonographic patterns of thyroid calcification in diagnosis of thyroid nodule.Methods 235 patients with 355 thyroid nodules were retrospectively analyzed.Nodule calcification size,distribution and pattern were observed.Results The incidence of calcification in benign and malignant nodules was 30.3％ (89/294)and 73.7％ (45/61) respectively.The difference had statistical significance(x2 =24.3,P ＜0.01).The rate of microcalcification in cancer was 47.5％ (29/61),higher than that in benign one 3.1％ (9/294)(x2 =99.1,P ＜ 0.01).Coarse calcification in benign nodules and malignant lesion was 27.2％ (80/294)and 26.2％ (16/61)respectively.The difference had no statistical significance(x2 =0.42,P ＞0.05).Conclusions Microcalcification of thyroid nodules is a specific index for thyroid carcinoma.Any type of sonographically detected calcification represents risk of malignancy.Not only microcalcification,these cases should raise the suspicion of malignancy in coarse calcification,especially involving a solitary nodule and irregular shape.

Objecfive To investigate the change of V-ATPase B subunits on epithelial to mesenchymal transition (EMT)in rat renal tubular epithelial cells (NRK52E) stimulated by transforming growth factor β1 (TGF-β1). Methods NRK52E cells were stimulated by TGF-β1 (10 μg/L)for O h(control),12 h,24 h,48 h,72 h after sefrum-free culture for 24 h.The mRNA and protein expression of E-cadherin,α-SMA,B2 and B1 subunits of V-ATPase were detected by real-time PCR,Western blotting and immunofluorescence. Results After stimulated by TGF-β1 (10 μg/L)for 48 h,the expression of α-SMA was markedly increased(P<0.05),but the expression of E-cadherin was dramatically decreased(P<0.05).Meanwhile,the expressions of V-ATPase subunit B2 was significantly increased (P<0.05).However,the B1 subunit distributed rarely in NRK 52E cells,and did not increase after TGF-β1 stimulation.Double-label immunofluoerscence staining also showed that the V-ATPase B2 subunit was increased in the cytosol.tending to accumulate to the cell membrane after TGF-β1 stimulation. Conclusions The main isoform of V-ATPase distributed in NRK52E cells is B2 subunit.B2 subunit is increased alone with TGF-β1-induced EMT.It may suggest that V-ATPase B2 subunit may play a potential role in TGF-β1-induced tubular EMT and renal fibrosis.

Objective To explore the protective effects of geranylgeranylacetone (GGA) on acute renal failure tats induced by isehemia reperfusion (IR) and the possible mechanism. Methods GGA (400 mg/kg) was administered to induce overexpression of heat shock protein 72 (HSP72) in the kidney of Sprague-Dawley (SD) rats. IR model was generated by temporary clamping the left renal artery for 45 minutes followed by right nephrectomy and 24 h reperfusion. A sham-operated group was used as normal control. 24 h after reperfnsion, rats were sacrificed. Blood was collected for measurement of serum creatinine (Scr) and blood urea nitrogen ( BUN ). Paraffin-embedded sections of the kidney were stained with PAS. Histological changes due to tubular damage were quantitated as tubular damage score. TUNEL assay was used to detect the apoptosis, and Western-blot was used to detect the expression of XIAP. Results After renal IR, the increased level of BUN and Scr, the tubular injury and the apoptosis of renal tubular epithelial cells were observed (P＜0.01). At the same time, the decreased level of XIAP was observed (P＜ 0.01). Compared with the control groups, the level of HSP72 expression was up-regulated in oral administration of GGA group (P＜0.05). The expression levels of BUN and serum creatinine were significantly decreased after IR injury in pre-conditioned rats with over-expression of HSP72 (P＜ 0.01 ). Kidney morphology was better preserved in GGA group. Rats with over-expression of HSP72 also revealed reduction of apoptotic cells by TUNEL stain and XIAP degradation by Western blot (P＜0.05). Conclusion GGA attenuates renal IR injury at least in part through inhibiting tubular cell apoptosis by decreasing XAIP degradation and restoring XIAP protein level.

Objective To investigate the role of C-Jun N-terminal kinase (JNK) in epithelial mesenchymal transition (EMT) induced by transforming growth factor β1 (TGF-β1) in rat peritoneal mesothelial cells(RPMCs). Methods RPMCs were harvested from the peritoneum of male Sprague-Dawley rats, then cultured in DMEM/F12 medium with 15% (V/V) FBS. After stimulation with TGF-β1, the expression of a-smooth muscle actin (α-SMA), E-cadherin and collagen I were detected in RPMCs. In some groups, the ceils were pretreated with SP600125, a specific inhibitor of JNK, for 4 hours before incubation with TGF-β1. The protein expression of phosphorylated JNK was detected by Western blotting. The mRNA and protein expression ofα-SMA, E-cadherin and collagen I were examined with RT-PCR and Western blotting, respectively.The intracellular distribution and expression of α-SMA was determined by indirect immunofluorescence. Results TGF-β1 could significantly increase the expression of α-SMA and collagen I, and decrease the expression of E-cadherin in RPMCs. TGF-α1 could stimulate the expression of phosphorylated JNK at 5 minutes with the peak at 10 minutes (P<0.01). The addition of SP600125 effectively inhibited TGF-β1-induced high expression of α-SMA and collagen I (P<0.05), and prevented TGF-β1-induced down-regulation of E-cadherin expression in RPMCs (P<0.05). The indirect immunofluorescence showed that the expression of intracellular α-SMA in RPMCs stimulated by TGF-β1 for 48 h increased significantly, which could be inhibited by SP600125. Conclusions JNK regulates epithelial mesenchymal transition induced by TGF-β1 in rat peritoneal mesothelial cells. JNK inhibitor may be used as a novel therapeutic agent for peritoneal fibrosis.

Objective To investigate the effects of peptide-binding domain (PBD) of heat shock protein (HSP) 72 on epithelial to mesenchymal transition (EMT) in rat renal tubular epithelial cells.Methods The expressions of wild-type HSP72,mutant of HSP72 lacking peptide binding domain (HSP72-△PBD) and HSP72-PBD were induced by plasmid transfection.NRK-52E ceils were stimulated by TGF-β1 for 48 h.The expressions of α-smooth muscle actin (α-SMA),E-cadherin,HSP72 and Smad3/p-Smad3 were detected by Western blot and immunofluorescence.Results After NRK-52E cells were stimulated by TGF-β 1 (10 μg/L) for 48 h,the expression of α-SMA was increased and the protein level of E-cadherin was decreased.Western blotting and immunofluorescence showed that over-expression of both HSP72 and PBD inhibited TGF-β1-induced up-regulation of protein α-SMA expression,down-regulation of protein E-cadherin.However,overexpression of HSP72-△PBD did not change the protein level of E-cadherin and α-SMA.In addition,over-expression of HSP72 and PBD significantly inhibited the phosphorylation of Smad3.Conclusion Inhibition of Smad3 activation and EMT by HSP72 is associated with the function of PBD.

Objective To analyze the changes of clinical and pathological features in the patients of IgA nephropathy with anemia.Methods Four hundred and nine patients of IgA nephropathy diagnosed by renal biopsy were classified into two groups:IgA nephropathy with nonanemia (group 1) and IgA nephropathy with anemia (group 2).Changes were studied retrospectively between the groups.Results Serum hemoglobin level was correlated with the clinical parameters of IgA nephropathy.Companed to group 1,changes in group 2 were as followed:serum creatinine increased,eGFR decreased,proteinuria increased; the global sclerosis,segmental sclerosis,crescents and tubulointerstitial lesions worsened.The glomerular and tubulointerstitial lesions were negatively correlated with serum hemoglobin and eGFR,but positively correlated with serum uric acid and proteinuria (P＜0.05).Multivariate Logistic regression analysis revealed that anemia was an independent risk factor for the tubulointerstitial lesion.Conclusion Clinical feature and pathological damages in the patients of IgA nephropathy with anemia are more serious than those with non-anemia.

Objective: To investigate the status of iodine nutrition and prevalence of goiter among children at ages of 8 to 10 years in Zhejiang Province in 2021, so as to provide insights into the management of iodine deficiency in children. Methods : A total of 90 counties (districts) were sampled as field survey sites from 11 cities of Zhejiang Province using a multi-stage stratified random sampling method in 2021, and non-residential children at ages of 8 to 10 years in these sites were sampled as study subjects. Subjects' household edible salt samples and random urine samples were collected. The iodine content in salt and urinary iodine level were determined using the direct titration method and arsenic-cerium catalytic spectrophotometry for evaluation of iodine nutrition among children. In addition, the lateral lobe of the thyroid gland was measured using ultrasound, and the prevalence of goiter was estimated. Results: A total of 19 363 children were recruited, including 9 710 male children and 9 653 female children, with a male to female ratio of 1.01︰1, and there were 10 704 urban children (55.28%) and 8 659 rural children (44.72%), 9 149 children living in coastal areas (47.25%) and 10 214 children in inland regions (52.75%). The mean iodine content was (19.79±9.25) mg/kg in salt, and the coverage of qualified iodized salt (15 252) was 78.77%. The coverage of qualified iodized salt was significantly lower in urban children than in rural children (76.70% vs. 81.21%; χ2=68.301, P<0.001), and was lower in children living in coastal regions than in inland regions (68.05% vs. 88.27%; χ2=1 270.769, P<0.001). The median urinary iodine concentration was 195.1 μg/L, and the household iodine content in salt correlated positively with urinary iodine concentration in children (rs=0.383, P<0.001). There were 2 885 children with iodine deficiency (14.90%), 7 137 children with adequate iodine (36.86%), 5 414 children with excessive iodine intake (27.96%), and 3 927 children with iodine overdose (20.28%), and the distribution of iodine nutrition in children varied significantly in regions (χ2=283.277, P<0.001) and gender (χ2=126.349, P<0.001). The prevalence of goiter was 2.45% among 7 195 children receiving ultrasound examinations, and a higher prevalence rate of goiter was detected in urban children than in rural children (2.76% vs. 2.00%; χ2=3.962, P=0.047). Conclusions The overall urinary iodine nutrition was adequate among children at ages of 8 to 10 years in Zhejiang Province in 2021, and the prevalence of goiter in children fell within the threshold defined in the criteria of elimination of iodine deficiency. However, the supervision of the iodized salt quality remains to be improved and iodine deficiency control remains to be reinforced.

Objective To investigate the effects of hemodialysis (HD) and peritoneal dialysis (PD) on the complications and outcomes after renal transplantation. Methods Clinical data of 402 renal transplant recipients maintained on dialysis for more than 3 months were retrospectively studied and divided into 2 groups: HD group(n=303)and PD group(n=99). Among them, 345 recipients were followed up for an average of (30.2±15.2) months. The impact of HD and PD on the acute rejection, delayed graft function (DGF), infection, chronic rejection and the graft and patient survival rates were analyzed. Results The mean dialysis duration was significantly longer in PD group and the hepatitis B infection rate was significantly higher in HD group. There were no signiticant differences between the HD and PD groups in regarding to primary disease for end-stage renal disease, age, gender, blood pressure, hemoglobin, HLA match, hot and cold ischemia time, and hepatitis C vires infection. The incidence of DGF, acute and chronic rejection, and cytomegalovirus and other infections between HD and PD groups were not significantly different. However, the graft loss happened more frequently in hepatatis B patients than that in non hepatitis B patients (19.23% vs 8.86%, P=0.021), and the post-transplant infection ocurred less in non hepatits B patients with PD. The acute rejection episodes were higher in HD patients who received pretransplant dialysis for more than 12 months (P<0.05). The overall recipients survival rates of HD and PD groups were similar (1-year: HD 94.34%, PD 91.25%;5-year: HD 92.83%, PD 90%), and the same as the graft survival rates in HD and PD groups (1-year: HD 93.21%, PD 96.25%;5-year: HD 87.17%, PD 91.25%). Conclusions The influences of PD and HD on the complications after renal transplantaton, 1-year and S-year recipients and graft survival rates are similar, so both HD and PD can be chosen as the pretransplant dialysis modality. As the incidence of acute rejection increases with time in HD, it is better to shorten the time of pretransplant dialysis to decrease the complication.

Objective To investigate the role of Notch signaling in the progression of peritoneal fibrosis in a rat model induced by high glucose dialysate. Methods Male Sprague Dawley rats were subjected to daily peritoneal dialysis (PD) with a lactate-buffered solution containing 4.25% glucose. They were sacrificed at 2 and 4 weeks after PD. The parietal thickness was measured with Masson staining. The expression of TGF-β1, E-cadherin, α-SMA and collagen Ⅰ was examined by immunoblotting. The expression of Notch ligand Jagged-1 and the negative Notch signaling regulato--Numb was analyzed by both immunoblotting and RT-PCR. The expression of a Notch nuclear target gene Hcs-1 was examined by RT-PCR. Results Both HE and Masson trichrome staining revealed an increase in peritoneal thickness with a loss of mesothelial cells and a rich of collagen matrix deposition in the submesothelial zone was evident at 4 weeks after PD. Meanwhile, compared to healthy rats, the expression of TGF-β1, ct-SMA and collagen Ⅰ was significantly increased, but the expression of E-cadherin was decreased in peritoneum after PD treatment. It was difficult to detect the Jagged-1 and Hes-1 expression in normal peritoneum, but their expression was graduaUy increased after PD. In contrast, the expression level of Numb, a negative regulator of Notch signaling, was dramatically decreased after PD. Conclusions Notch signaling is activated during the process of PD-induced peritoneal fibrosis and the activation of Notch signaling is associated with the loss of negative regulation of Notch signaling via decreased expression of Numb. Inhibition of Notch signaling via overexpression of its negative regulators such as Numb may be a novel therapeutic approach for peritoneal fibrosis in PD patients.