Objective To explore the possible role of vitamin D in the pathogenesis of IgA nephropathy (IgAN). Me-thods After the IgAN model was successfully induced at 12 weeks, the BALB/C mice were randomly divided into IgAN group (n=15) and IgAN+VitD group (n=15). The nephrosis mice were administrated with 100 μl/d propylene glycol or propyl-ene glycol+1,25(OH)2D, 3 ng/(100g?d), for 6 weeks. The control group was setted (n=15). The level of 24 hour urine protein was determined at week 0, 12 and 18. At week 18, the levels of serum 25(OH)D, ifbroblast growth factor 23 (FGF23) and galactose-deifcient IgA1 (Gd-IgA1) were detected. The mRNA and protein expressions of interleukin-21 (IL-21) in Peyer’s patches (PPs) were detected by lfuorescent quantitative reverse transcription-polymerase chain reaction and western blot respectively. The protein expression of Bcl-6 was detected by western blot. The percentages of Tfh cells/T lymphocytes, B220+IgM+/B lympho-cytes, B220+IgA+/B lymphocytes, B220-IgA+/B lymphocytes in PPs were determined by lfow cytometry. Results Compared with control group, the levels of 24 hour urine protein, FGF23 and Gd-IgA1 were increased, serum 25(OH)D was decreased, the mRNA and protein expressions of IL-21 and the protein level of Bcl-6 were increased, the percentages of Tfh cells/T lym-phocytes, B220+IgM+/B lymphocytes, B220+IgA+/B lymphocytes, B220-IgA+/B lymphocytes were elevated in IgAN group (P<0.05). These indicators were improved in IgAN+VitD group. Compared with the IgAN group, the differences were statisti-cally signiifcant (P<0.05), however compared with control group, some indicators showed no signiifcant differences (P>0.05). Conclusions 1,25(OH)2D may protect the microenvironment of PPs in IgAN through inhibiting the differentiation of Tfh cells and B cells and the generation of Gd-IgA1.

Objective:To investigate the baseline levels of 25-hydroxy-vitamin D [25-(OH) D] in 3340 hospitalized children, analyze the 25-( OH) D levels in children with different diseases, age and gender in different seasons, and study the correlation between the 25-( OH) D levels and the clinical indicators. Methods:Totally 3340 hospitalized children were randomly selected, the 25-( OH) D levels were detected by an ELISA method, and Pearson correlation analysis of 25-( OH) D levels and clinical indicators such as liver function, myocardial enzymes, immunoglobulins, lymphocyte subtypes and thyroid function was performed. Results: The serum 25-(OH) D level in 3340 cases (1850 male, 1490 female) was (33. 00 ± 13. 42) ng·m1 -1, and that in those with neonatal diseases was the lowest followed by those with primary nephrotic syndrome, Henoch-schordeinpurpura and juvenile idiopathic arthritis. The ser-um 25-( OH) D levels in the premature was higher than that in full term infant. Except the newborn, the level of serum 25-( OH) D gradually reduced along with the age increase, while the percentage of insufficiency gradually increased. The serum 25-( OH) D level between the male and the female had no significant difference. The 25-( OH) D levels of hospitalized children were the highest in sum-mer. The serum level of 25-( OH) D was positively correlated with body mass index ( BMI) , alanine transaminase ( ALT) , aspartate transaminase (AST), creatine kinase (CK), creatine-phosphokinase (CK-MB), lactate dehydrogenase(LDH), free T4 (FT4) and free T3 (FT3), and negative correlation with alkaline phosphatase(ALP), immunoglobulin E (IgE) and immunoglobulin M (IgM). Conclusion:The incidence of vitamin D insufficiency is high in hospitalized children. The level of vitamin D is different among various diseases. The level of serum 25-( OH) D may have certain relevance with BMI, allergies, myocardial damage and thyroid function.

Objectives To study the protective effects of vitamin D (VitD) on podocyte insulin resistance and its mecha-nisms. Methods Immortalized mouse podocytes in vitro were randomly divided into 4 groups:podocytes+5 mmol/L glucose group (group A);podocytes+5 mmol/L glucose+1 nmol/L propylene glycol group (group B);podocytes+30 mmol/L glucose+1 nmol/L propylene glycol group (group C); podocytes+30 mmol/L glucose+1 nmol/L propylene glycol+1 nmol/L VitD group (group D). The percentage of podocyte apoptosis was determined after 48 h of incubation. Podocyte viability was assessed by MTT assay. The mRNA expressions of vitamin D receptor (VDR) and insulin receptor substrate protein 1 (IRS1) in podocyte were detected by RT-PCR. Western blot analysis was performed to measure the protein levels of p-IRS1/IRS, p-Akt/Akt and p-ERK1/2/ERK1/2. Results There were significant differences in apoptosis percentage, viability and the expression of VDR, IRS1, p-ERK1/2 of podoctyes(P<0.05)among 4 groups. There was no difference in p-Akt/Akt expression among 4 groups(P>0.05). Compared with group A, B , and D, the percentage of podocyte apoptosis in group C was significantly increased, the cell viabi-lity was decreased, the expressions of VDR and IRS1 mRNA and p-IRS1 and p-Akt proteins were down-regulated, whereas p-ERK1/2 was up-regulated in group C. The levels of p-IRS1/IRS1, p-Akt/Akt, p-ERK1/2/ERK1/2 had no statistical differences in group A, B, and D (P>0.05). Conclusions VitD-VDR system alleviates podocyte apoptosis induced by high glucose, and acti-vates insulin signaling pathway and counteracts insulin resistance signal to improve podocyte insulin resistance.

AIM: To investigate whether glucosylceramide synthase (GCS) regulates apoptosis-related gene bcl-2 expression via MEK/ERK signaling pathway , thus enhancing drug resistance of K 562/A02 human leukemia multidrug resistant cell line.METHODS:siRNA targeting GCS was transfected into K562/A02 cells.Bcl-2, p-ERK and total ERK expression at mRNA and protein levels after GCS knockdown were detected by real-time PCR and Western blotting .After exposed to MEK-ERK pathway inhibitor U0126, the expression of Bcl-2 at mRNA and protein levels also was analyzed by real-time PCR and Western blotting , respectively.The viability of the cells was evaluated by CCK-8 assay.RESULTS:The expression of GCS and Bcl-2, as well as MEK/ERK signaling were significantly inhibited in K 562/A02 cells by GCS siRNA transfection compared with negative control group .Inactivation of MEK/ERK signaling due to U0126 treatment de-creased Bcl-2 mRNA and protein levels in a concentration-dependent manner , and sensitized K562/A02 cells to adriamy-cin.CONCLUSION:GCS may affect the expression of apoptosis-related gene bcl-2 by MEK/ERK signaling pathway , thus regulating multidrug resistance of human leukemia K 562/A02 cells.

Objective To analyze the distribution and drug resistance of pathogens for bacterial infection after lung transplantation,so as to provide evidence for clinical prophylactic strategies postoperation and reasonable use of antibiotics.Method The bacterial distribution and drug resistance of 81 recipients after lung transplantation in our hospital were retrospectively analyzed from May 2009 to October 2012.The VITEK-32 full-automatic microbial identification system (Biomerieux,France)and its supplementary reagent were used for bacterial identification and drug sensitive test.The data were statistically analyzed by using the software SPSS 13.0.Result There were 67 cases of bacterial infection in the 81 recipients after lung transplantation and the infection rate was 82.72％ (67/81).The infection was caused by one kind of bacteria in 20 patients,two kinds of bacteria in 23 patients and multiple bacteria in 24 patients.157 strains pathogenic bacteria were produced,and the grampositive bacilli and the gram-negative bacilli accounted for 12.74％ and 87.26％ respectively.The most common pathogens for the bacterial infection were Acinetobacter baumannii,Klebsiella pneumoniae,Pseudomonas aeruginosa,Stenotrophomonasmaltophilia,Escherichia coli and Staphylococcus aureus.Most of the bacterial infections occurred in the early period (≤1 month) after lung transplantation and most non-fermentative bacterial pathogens were resistant to multi-antibiotics.Conclusion The bacterial infection rate is high after lung transplantation.The rational use of antibiotics in clinical practice should be adjusted according to the bacterial distribution and drug resistance.

OBJECTIVETo explore the effect of organophosphorus insecticides (OPs) on G protein-coupled receptor kinase 2 mediated phosphorylation of M2 muscarinic receptors in vitro and to understand an alternative target of the OPs for human and other animals.

METHODSThe acetylcholine M2 muscarinic receptors (mAChR2) were purified from rat brain by single step affinity chromatography. In vitro experiments, the purified mAChR2, G-protein coupled receptor kinase 2 (GRK2) and the (gamma-p32) labeled ATP were incubated with paraoxon (PO), chlorpyrifos oxon (CPO) or chlorpyrifos (CPF) of varying concentrations. The proteins were separated by the polyacrylamide gel electrophoresis. The gels were dried and the phosphorylation of mAChR2 was detected with autoradiograms. Bands containing M2 receptor were excised and counted by liquid scintillation.

RESULTSCPO inhibited phosphorylation of M2 muscarinic receptors by GRK2 with a median inhibition concentration (IC(50)) at 70 micromol/L. CPF also inhibited M2 receptors phosphorylation, but was less potent and less efficacious than that of CPO. PO and parathion (PT) had little effect on the receptor phosphorylation under the same conditions. CPO and CPF didn't inhibit the beta2 Adrenalin (beta2-AR) receptor phosphorylation also mediated by GRK2.

CONCLUSIONCPO and CPF can selectively inhibit the GRK2 mediated mAChR2 phosphorylation while PO and PT have no this effect.

Animals ; Chlorpyrifos ; analogs & derivatives ; toxicity ; Cholinesterase Inhibitors ; toxicity ; G-Protein-Coupled Receptor Kinase 2 ; Paraoxon ; toxicity ; Phosphorylation ; Rats ; Receptor, Muscarinic M2 ; metabolism ; beta-Adrenergic Receptor Kinases ; metabolism ; physiology

OBJECTIVETo evaluate the sulcus depth changes of bilateral alveolar clefts after bone grafting with different techniques.

METHODSForty-two patients with bilateral alveolar clefts were treated in the Department of Oral and Maxillofacial Surgery of Xiangya Hospital, Central South University, including three bilateral alveolar clefts with bilateral cleft lips and thirty-nine bilateral alveolar clefts with bilateral complete cleft lip and cleft palate. Three techniques were used to repair the bilateral alveolar clefts, named: bucket-handle mucosal flap, bilateral buccal mucosal flaps and bilateral buccal mucosal flaps with premaxillary separation and bone grafting. Postoperatively, the depth of the sulcus was evaluated with the following four grades: 0) no change, I) one-third shallow, II) two-thirds shallow, III) disappear.

RESULTSForty-two bilateral alveolar clefts were repaired with the above mentioned techniques with the results: Grade 0 in 20 patients (47.6%), grade I in 10 patients (23.8%), grade II in 10 patients (23.8%), grade III in 2 patients (4.7%). In 6 patients with the bucket-handle mucosal flap technique and the technique of the bilateral buccal mucosal flaps with the premaxillary bone graft, the depth of the sulcus was reduced to two-thirds in 4 patients (66.7%), and the sulcus totally disappeared in 2 patients (33.3%).

CONCLUSIONSThe above mentioned techniques could reduce the depth of the sulcus, therefore, the surgeons should notice this complication and prevent it during the operation.

Adolescent ; Adult ; Bone Transplantation ; Child ; Cleft Palate ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps ; Treatment Outcome

OBJECTIVEFragile X syndrome (FXS) may be identified by many methods, such as PCR assay and Southern blot. However, each method has its limits or shortcomings. This study explored the reliability of the rapid, convenient and inexpensive hair root fragile X mental retardation protein (FMRP ) assay in the identification of FXS.

METHODSFMRP in hair roots was determined by immunohistochemistry assay in 80 healthy children, in 40 children with mental retardation of unknown etiology and in 12 family members in one pedigree of FXS. FXS was confirmed by 7-deza-dGTP PCR.

RESULTSThere was a high expression of FMRP in hair roots (> or =80%) in healthy children. Two children were confirmed with FXS by 7-deza-dGTP PCR in 40 children with mental retardation of unknown etiology. FMRP expression was 10% and zero respectively in the two children. The other 38 children had FMRP expression of more than 80%. FMRP was not expressed in the two cases of FXS from the pedigree of FXS.

CONCLUSIONSInexpensive, rapid and convenient hair root FMRP assay is reliable for the diagnosis of FXS and may be widely applied for screening and diagnosing FXS in children with mental retardation.

Adolescent ; Child ; Child, Preschool ; Female ; Fragile X Mental Retardation Protein ; analysis ; Fragile X Syndrome ; diagnosis ; genetics ; Hair ; chemistry ; Humans ; Infant ; Male ; Polymerase Chain Reaction

OBJECTIVETo define the differences in gene expression patterns between glycidyl methacrylate (GMA)-transformed human lung fibroblast cells (2BS cells) and controls.

METHODSThe mRNA differential display polymerase chain reaction (DD-PCR) technique was used. cDNAs were synthesized by reverse transcription and amplified by PCR using 30 primer combinations. After being screened by dot blot analysis, differentially expressed cDNAs were cloned, sequenced and confirmed by Northern blot analysis.

RESULTSEighteen differentially expressed cDNAs were cloned and sequenced, of which 17 were highly homologous to known genes (homology = 89%-100%) and one was an unknown gene. Northern blot analysis confirmed that eight genes encoding human zinc finger protein 217 (ZNF217), mixed-lineage kinase 3 (MLK-3), ribosomal protein (RP) L15, RPL41, RPS 16, TBX3, stanniocalcin 2 (STC2) and mouse ubiquitin conjugating enzyme (UBC), respectively, were up-regulated, and three genes including human transforming growth factor beta inducible gene (Betaig-h3), alpha-1,2-mannosidase 1A2 (MAN 1A2) gene and an unknown gene were down-regulated in the GMA-transformed cells.

CONCLUSIONAnalysis of the potential function of these genes suggest that they may be possibly linked to a variety of cellular processes such as transcription, signal transduction, protein synthesis and growth, and that their differential expression could contribute to the GMA-induced neoplastic transformation.

Air Pollutants, Occupational ; toxicity ; Carcinoma, Squamous Cell ; genetics ; pathology ; Cell Line, Transformed ; Epoxy Compounds ; toxicity ; Fibroblasts ; cytology ; drug effects ; Gene Expression Profiling ; Glycoproteins ; metabolism ; Humans ; Lung ; cytology ; Male ; Mannosidases ; drug effects ; metabolism ; Methacrylates ; toxicity ; Mitogen-Activated Protein Kinase 3 ; drug effects ; metabolism ; Oligonucleotide Array Sequence Analysis ; Ribosomal Proteins ; metabolism ; Signal Transduction ; genetics ; Transforming Growth Factor beta ; drug effects ; metabolism ; Ubiquitins ; metabolism ; Zinc Fingers ; drug effects ; physiology

OBJECTIVETo evaluate the genotoxic and nongenotoxic effects of short-term exposure to glycidyl mathacrylate (GMA) on human lung fibroblast cells (2BS cells) in vitro.

METHODSDNA strand breakage was determined by single cell gel electrophoresis, and DNA ladder formation assay and flow cytometric analysis were carried out to detect apoptic responses of cells to GMA exposure. The HPRT gene mutation assay was used to evaluate the mutagenicity, and the effect of GMA on gap junctional intercellular communication (GJIC) in the exposed cells was examined with the scrape loading/dye transfer technique. The ability of GMA to transform 2BS cells was also tested by an in vitro cell transformation assay.

RESULTSExposure to GMA resulted in a dose-dependent increase in DNA strand breaks but not apoptic responses. GMA was also shown to significantly induce HPRT gene mutations and morphological transformation in 2BS cells in vitro. In contrast, GMA produced a concentration-dependent inhibition of GJIC.

CONCLUSIONSGMA elicits both genotoxic and nongenotoxic effects on 2BS cells in vitro. The induction of DNA damage and gene mutations and inhibition of GJIC by GMA may casually contribute to GMA-induced cell transformation.

Cell Communication ; Cell Differentiation ; Comet Assay ; DNA Damage ; DNA Mutational Analysis ; Epoxy Compounds ; toxicity ; Fibroblasts ; Gap Junctions ; Humans ; Hypoxanthine Phosphoribosyltransferase ; genetics ; Lung ; cytology ; Methacrylates ; toxicity