AIM: To investigate the expressions of the cyclin dependent kinase inhibitor (CKI) in human corneal epithelium.METHODS: The expressions of CKI, P27, P21 and proliferating cell nuclear antigen (PCNA) were tested in different regions of corneal epithelium by SP immunohistochemistry.RESULTS: Limbal basal cells stained positively for PCNA while central corneal epithelium cells stained negatively for PCNA, their difference was statistically significant (P＜0.01).Positive staining for P27 and P21 were observed in central epithelium, but there was no positive staining in limbal epithelium. Their difference were also statistically significant (P＜0.01).CONCLUSION: The different expressions of CKI P27, P21and PCNA in different corneal epithelial regions suggest that in limbal basal layer there are a group of cells that have higher proliferative capacity staying in G1 status, namely stem cell.

Objective To observe the expression of bone morphogenetic protein 7 (BMP-7) in different stages of diabetic rats and investigate the potential role of BMP-7 in diabetic nephropathy (DN). Methods Wistar rats were divided into diabetes mellitus(DM) group induced by intravenous injection of streptozotocin (65 mg/kg) , and normal control group injected with citrate buffer. The rats were sacrificed at day 30,60,90,120,150 and 180 following injection, respectively. The blood glucose,Scr, urinary albumin and urinary creatinine were measured in each rat before sacrifice. The renal pathological changes were examined with hematoxylin and eosin (HE),periodic acid-silver-methenamine staining (PASM) and protein expression of collagenⅣ(ColⅣ). The mRNA and protein expression of BMP-7 and TGF-?1 in kidney were detected by reverse transcription-polymerase chain reaction (RT-RCR) and immunohistochemical staining respectively, and were quantified by computer image analysis system. Results The levels of blood glucose and urinary albumin in diabetic group were remarkably higher than those in control group (P

Objective To investigate the respiratory mechanics and treatment outcomes of different types of mechanical ventilation for patients with neonatal acute respiratory distress syndrome(ARDS):the high frequency oscillation ventilation + pulmonary surfactant (HFOV+PS),conventional mechanical ventilation + pulmonary surfactant(CMV+PS),conventional mechanical ventilation(CMV). Methods Seventy-five cases with neonatal ARDS,25 cases in the HFOV+PS group,30 cases in the CMV+PS group,20 cases in the CMV group. Patients in the former two groups received 70 mg/kg PS at a time. PaO2,PaCO2,PaO2/FiO2,oxygenation index(OI)and respiratory index(RI)were detected at 0 h,12 h,24 h,48 h,72 h post-mechanical ventilation. Results At 12, 24,and 48 hours post-mechanical ventilation,patients in the HFOV+PS group had a significantly higher level of PaO2 and a significantly lower level of PaCO2 than those of patients in the CMV+PS group and the CMV group(P<0.05). At 12,24,48,and 72 hours post-mechanical ventilation,patients in the HFOV+PS group had a significantly higher level of PaO2/FiO2 and significantly lower level of OI and RI than those of patients in the CMV+PS group and the CMV group (P < 0.05,respectively). Patients in the HFOV+PS group also had significantly shorter durations of mechanical ventilation and oxygen usage than those of patients in the CMV+PS group and the CMV group (P < 0.05). No significant differences were observed in Gas leak,the incidence of intracerebral haemorrhage and cure rate among the three groups. Conclusions Application of HFOV with PS therapy for patients with neonatal ARDS can timely improve the oxygenation,shorten the time of mechanical ventilation and the usage of oxygen,without increasing complications.

Objective To investigate the effects of high-flux hemodialysis (HFD) on fibroblast growth factor-23 (FGF-23) levels in maintenance hemodialysis (MHD) patients and its clinical significance.Methods Sixty uremia patients were divided into HFD group and hemodialysis (HD)group and observed for 12 months.Flow mediated dilation (FMD),cardiac ultrasonography,levels of FGF-23,serum phosphorus,serum calcium,25-(OH)D3,parathyroid hormone (PTH),homocysteine (Hcy) and interleukin-6 (IL-6) were tested in all patients before and after treatment.The correlation of above indexes were analyzed.Results No statistical difference were found in primary disease,age and duration of dialysis in two groups.The levels of FGF-23 [(56.07±26.63) vs (85.53±40.54) ng/L,P ＜0.01],IL-6 [(3.37±2.48) vs (5.59±2.53) ng/L,P ＜ 0.05] and Hcy [(21.13±6.95) vs (29.40±11.66) μmol/L,P ＜ 0.05] decreased and FMD,25-(OH)D3 [(27.3± 10.26) vs (23.15± 10.73) μg/L,P ＜ 0.05] increased significantly after the treatment of HFD.There were no significant changes in the HD group.The baseline FMD was negatively correlated with FGF-23 (r =-0.413,P ＜ 0.05) and Hcy (r =-0.301,P ＜0.05).The baseline LVMI was correlated with FGF-23 (r =0.464 P ＜ 0.05).After one year's trearmeat of HFD,the changes of FMD(/△ FMD) was negatively correlated with the changes of FGF-23 (/△ FGF-23)(r =-0.347,P ＜ 0.05).Conclusions HFD can improve FMD and decrease FGF-23 levels.The improvement of FMD may be related to the decreased level of FGF-23.The effect of FGF-23 on FMD should be independent of serum phosphate.

This study is to investigate the effect of artesunate on transforming growth factor-beta1 (TGF-beta1) induced epithelial-mesenchymal transition (EMT) and its possible mechanism. After the in vitro cultured RLE-6TN cells were treated with TGF-beta1 then artesunate intervened on it, after 24 h, expression of the markers of mesenchymal cell was assayed using Western blotting and real-time PCR analysis. Western blotting was also used to detect the effect of TGF-beta1 on the Smad3 and Smad7 expressions of RLE-6TN cells. Morphological alterations were examined by phase-contrast microscope, and ultrastructure changes by electron microscope. Incubation of RLE-6TN cells with TGF-beta1 resulted in the up-regulation of the expression of the mesenchymal cell markers, after artesunate intervened on it, resulted in the down-regulation of the expression. Meanwhile, incubation with artesunate intervened on RLE-6TN cells could lead to the apparent down-regulation of the expression of Smad3 and up-regulation of Samd7 and the transition of RLE-6TN cells to mesenchymal-like by TGF-beta1 induction, after artesunate intervened on it, RLE-6TN cells to epithelial-like. TGF-beta1 induced epithelial-mesenchymal transition process; artesunate can inhibit TGF-beta1-induced epithelial-mesenchymal transition process, the possible mechanism is up-regulation of the expression of Smad7 and down-regulation of the expression of Smad3, meanwhile inhibits phosphorylation of Smad3.
Actins ; genetics ; metabolism ; Animals ; Artemisia ; chemistry ; Artemisinins ; isolation & purification ; pharmacology ; Cell Line ; Cell Proliferation ; drug effects ; Epithelial Cells ; cytology ; metabolism ; Epithelial-Mesenchymal Transition ; drug effects ; Idiopathic Pulmonary Fibrosis ; pathology ; Plants, Medicinal ; chemistry ; Pulmonary Alveoli ; cytology ; RNA, Messenger ; metabolism ; Rats ; Smad3 Protein ; genetics ; metabolism ; Smad7 Protein ; genetics ; metabolism ; Transforming Growth Factor beta1 ; pharmacology ; Vimentin ; genetics ; metabolism

Proteins extracted from two varieties of Chinese roses leaves were separated by two- dimensional polyacrylamide gel electrophoresis (2-DE) with immobilized pH gradient (IPG). Many difference proteins were isolated with molecular weights ranging 10-30 kDa and pI5-6. Three proteins of high levels observed in a gel were excised and identified using peptide mass fingerprinting and MS-MS. A summary of the identified proteins and their putative functions are presented. They are identified as eIF-5A、LEA protein and Hsp17. 5. Functions of these proteins in plant tolerance to high temperature were discussed.

Objective To detect the levels of 25-hydroxyvitamin D3 and analyse the related clinical factors in patients with chronic kidney disease (CKD).Methods Patients with CKD in our department from March 1st to July 1st were enrolled continuously.The level of 25-hydroxyvitamin D3 and intact parathyroid hormone(iPTH) were detected by electrochemiluminescence and immunochemiluminescent respectively.Serum calcium,phosphorus and albumin were measured by automatic biochemical instrument.Results 127 patients were selected and the average age was (60.9?15.3).The mean level of 25-hydroxyvitamin D3 was (12.06?6.41)?g/L.82.6% patients had vitamin D deficiency and 96.9% patients had vitamin D insufficiency.The level of 25-hydroxyvitamin had no statistics difference D3 between stage 1,2 and 3 CKD patients but was much hihger than that of stage 4 and non-haemodialysis stage 5 patients.The level of 25-hydroxyvitamin D3 was not related to serum calcium,phosphorus and iPTH,while positively related to albumin and eGFR and negatively related to serum creatinine and total cholesterol.Conclusion Vitamin D deficiency and insufficiency are frequent in CKD patients and deteriorate with the progress of kidney function impairment.The level of 25-hydroxyvitamin D3 is not related to the traditional CKD-MBD index such as serum calcium,phosphorus and iPTH.

OBJECTIVETo compare the early clinical effects of Activ C cervical disc replacement (ACDR) and anterior cervical discectomy and fusion (ACDF) in treating single-level cervical spondylosis.

METHODSThe clinical data of 76 patients with single-level cervical spondylosis underwent surgery from July 2009 to September 2012 were retrospectively analyzed. Among them, 28 patients were treated with ACDR (ACDR group), including 18 males and 10 females, aged from 32 to 62 years old with an average of (45.2±6.2) years; and 48 patients were treated with ACDF (ACDF group), including 28 males and 20 females, aged from 33 to 60 years old with an average of (45.8±6.4) years. Visual analogue scale (VAS), Japanese Orthopedics Association (JOA) score, Short Form-36 (SF-36), imaging data were used to assess the clinical effects after operation.

RESULTSA total of 76 patients were followed up from 6 to 24 months with an average of 13.2 months. VAS of neck pain and brachialgia were improved in all patients after operation (P<0.05), there was no significant difference between two group (P>0.05). Somato-score and psycho-score of SF-36 of two groups were obviously increased (P<0.05), ACDR group was better than that of ACDF group (P<0.05). In ACDR group, there was no significant difference in the range of motion of surgical segments and adjacent segments between preoperative and postoperative (P>0.05); heterotopic ossification around the edge of vertebral body occurred in 1 case on the 6th month after operation, no fusion was found on the 1st year after operation. In ACDF group, the adjacent vertebral disease occurred in 1 case and the patient underwent the reoperation.

CONCLUSIONActiv C cervical disc replacement can reduce the degeneration of adjacent segments and its early outcomes for the treatment of single-level cervical spondylosis are satisfactory, but the long-term effects still need study.

Adult ; Cervical Vertebrae ; surgery ; Diskectomy ; methods ; Female ; Humans ; Male ; Middle Aged ; Spinal Fusion ; methods ; Spondylosis ; surgery ; Total Disc Replacement ; methods

Objective To explore whether the change of S phase kinase-associated protein 2 (Skp2) expression could regulate mesangial cell proliferation. Methods Skp2 siRNA and control siRNA, pIRES-GFP-Skp2 plasmid and pIRES-GFP plasmid were designed and synthesized. Cell transfection was performed by Lipofectamine 2000. Skp2 mRNA and protein levels were detected by semiquantitative PCR and Western blotting respectively. Primary culture rat mesangial cells were divided into 6 groups: 0%FCS, 20%FCS, 10%FCS+pIRES-GFP plasmid, 10%FCS+pIRES-GFP-Skp2 plasmid, 20%FCS+control siRNA, 20%FCS+Skp2 siRNA. Cell number was detected by MTT. S phase entry was measured by BrdU labeling. Cell cycle profile was determined by flow cytometric analysis. Results Skp2 mRNA level was significantly down-regulated by Skp2 siRNA compared to control siRNA. Skp2 protein level increased after pIRES-GFP-Skp2plasmid transfection compared to pIRES-GFP plasmid. MTT, BrdU labeling and cell cycle profile demonstrated that cell number (A: 0.419±0.088 vs 0.305±0.036, P＜0.01) and S-phase cells (BrdU labeling positive cell: 0.21±0.04 vs 0.15±0.03, P＜0.01;S-phase cell number:20.18±0.64vs 14.33±0.37, P＜0.01) obviously increased after Skp2 plasmid transfection, while decreased after Skp2 siRNA transfection compared to control groups (A: 0.328±0.069 vs 0.482±0.133, P＜0.01;BrdU labeling positive cell: 0.17±0.01 vs 0.24±0.00, P＜0.01; S-phase cell number: 16.52±0.75vs 23.81 ±1.25, P＜0.01). Conclusion Over-expression of Skp2 stimulates mesangial cell proliferation while down-regulated expression of Skp2 inhibits mesangial cell proliferation.

Objective To investigate the effects of IGF-Ⅱ on adriamycin-induced apoptosis of hepatocellular carcinoma cell line(HepG2).Methods Cultured HepG2 cells were divided into:①group A:control group;②group B:ADM group(200 ng/ml);③group C:2 ng/ml IGF-Ⅱ+200 ns/ml ADM group;④group D:20 ng/ml IGF-Ⅱ+200 ng/ml ADM group;⑤group E:200 ng/ml IGF-Ⅱ+200 ng/ml ADM group. After HepG2 were treated for 48 h,MTT colorimetry was performed to determine the cell viability,Flow cytometry was used to detect the cell apoptosis rate,and Weastern-blot was performed to evaluate the expression of survivin. Results The cell viability of group A、B、C、D、E was respectively 0.568±0.025、0.201±0.020、0.232±0.027、0.268±0.013 and 0.304±0.019;The cell apoptosis rate of group A、B、C、D;E was respectively 6.9％±1.3％、35.4％±2.1％、31.2％±2.2％、26.4％±1.7％and 21.7％±1.9％;Survivinβ-actin of group A、B、C、D、E was respectively 0.527±0.039、0.147±0.081、0.311±0.069、0.421±0.033 and 0.469±0.031.The cell viability in IGF-Ⅱ+ADM group was better than ADM group(P＜0.01),the cell apoptosis rate in IGF-Ⅱ+ADM group was significantly lower than ADM group(P＜0.01),the expression level of survivin in IGF-Ⅱ +ADM group was significantly higher than ADM group(P＜0.01). Conclusions IGF-Ⅱ inhibits ADM-induced apoptosis of HepG2 cells,probably by up-regulating cell's survivin expression.