Adenoma ; metabolism ; pathology ; surgery ; Humans ; Keratin-7 ; metabolism ; Kidney Neoplasms ; metabolism ; pathology ; surgery ; Kidney Transplantation ; Living Donors ; Male ; Mucin-1 ; metabolism ; Young Adult

Objective To investigate the effects of recombinant human fibronectin fragment (RetroNectin) combined with anti-CD3 monoclonal antibody (CD3Ab) on the proliferation and cytotoxicity of cytokine-induced killer cells (CIK) from acute leukemia (AL). Methods Mononuclear cells (MNCs) were isolated from peripheral blood of complete remission AL pa-tients. The MNCs were cultured in vitro by precoating with RetroNectin (RN group), CD3Ab (CD3Ab group), RetroNectin com-bined with CD3Ab (RN+CD3Ab group) and traditional method (control group) to generate CIK. The changes of growth rate, characterization, cytotoxicity and apoptosis of CIK were determined between groups. Results The amplification of CIK was higher in experimental group than that of control group, and the amplification of CIK was higher in group RN+CD3Ab than that of in group RN and group CD3Ab (P＜0.05). The expression of CD25 positive cells was higher in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P＜0.05).The percentage of G1 stage cells was lower in group RN and group RN+CD3Ab than that of group CD3Ab and control group. The percentage of S stage cells was higher in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P＜0.05). The cytotoxicity was higher in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P＜0.05) at the E/T scope 40∶1.The percentage of apoptotic cells was lower in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P ＜ 0.05). Conclusion These in vitro studies suggest that a higher activity of immune cells could be obtained by CIK cells cultured by precoating Ret-roNectin and CD3Ab.

Objective:To know the work status of clinical pharmacy in medical institutions of Guizhou province. Methods:Ques-tionnaires were used to analyze the situation of clinical pharmacy in 108 medical institutions of Guizhou province. Results: A total of 246 questionnaires were taken back, and among the 231 valid questionnaires were received including gradeⅡor above hospitals. The main contents of clinical pharmacy work carried out in medical institutions included 7 aspects: pharmacists ’ participation in ward rounds, which accounted for 47. 11%; pharmacists’ participation in case consultation, which accounted for 16. 65%; pharmacists’ participation in teaching practice, which accounted for 38. 84%; pharmacists’ participation in prescription evaluation and analysis, which accounted for 72. 73%;pharmacists’ participation in antimicrobial drug monitoring and drug use evaluation, which accounted for 62. 37%;pharmacists’ participation in drug counsultation and education, which accounted for 58. 68%;pharmacists’ participation in adverse drug reaction monitoring and supervision, which accounted for 77. 32%. Conclusion:The development of clinical pharmacy in Guizhou province still lags behind, and the number of clinical pharmacists is insufficient, which can’ t meet the growing demand for personalized medicine. In particular, the development of clinical pharmacy is restricted by the limited pharmaceutical service. The cog-nition degree of pharmacist group in Guizhou province has been improved. However, the number and the service quality of clinical pharmacists need to be improved further.

Animals ; Coculture Techniques ; Colonic Neoplasms ; pathology ; therapy ; Cytokines ; pharmacology ; Cytotoxicity, Immunologic ; Dendritic Cells ; immunology ; Female ; Immunophenotyping ; Immunotherapy, Adoptive ; Interferon-gamma ; biosynthesis ; Interleukin-12 ; biosynthesis ; Killer Cells, Natural ; immunology ; Lung Neoplasms ; prevention & control ; secondary ; Lymphocyte Activation ; Mice ; Mice, Inbred BALB C

BACKGROUNDMutations in fumarylacetoacetate hydrolase (FAH) gene can lead to tyrosinemia type 1 (HT1), a relatively rare autosomal recessive disorder. To date, no molecular genetic defects of HT1 in China have been described. We investigated a Chinese family with a HT1 child to identify mutations in FAH.

METHODSDNA sequencing was used for mutations screening in FAH gene. Real-time polymerase chain reaction (PCR) was performed to determine the FAH gene expression level. To confirm the presence of degradation by the nonsense-mediated mRNA decay pathway (NMD), the fragments containing R237X mutations were analyzed by primer introduced restriction analysis-polymerase chain reaction (PIRA-PCR) and cDNA sequencing. Finally, the effects of the mutations reported in this study were predicted by online softwares.

RESULTSA boy aged 3 years and 8 months was diagnosed clinically with HT1 based on his manifestations and biochemical abnormalities. Screening of FAH gene revealed two heterozygous mutations R237X and L375P transmitted from his mother and father respectively. In this pedigree, the amount of FAH mRNA relative to a healthy control was 0.44 for the patient, 0.77 for his mother and 1.07 for his father. Moreover, both PIRA-PCR and cDNA sequencing showed significant reduction of the FAH mRNA with R237X nonsense mutation. The missense mutation of L375P was not reported previously and prediction software showed that this mutation decreased the stability of protein structure and affected protein function.

CONCLUSIONSThis is the first case of HT1 analyzed by molecular genetics in China. The R237X mutation in FAH down- regulates the FAH gene expression, and the L375P mutation perhaps interrupts the secondary structure of FAH protein.

Child, Preschool ; China ; Humans ; Hydrolases ; genetics ; Male ; Molecular Sequence Data ; Mutation ; Mutation, Missense ; genetics ; Nonsense Mediated mRNA Decay ; genetics ; Real-Time Polymerase Chain Reaction ; Tyrosinemias ; genetics

OBJECTIVETo explore the association between genetic polymorphisms of TGF-beta (TGF-beta) and susceptibility to pneumoconiosis.

METHODSOne hundred and seventeen patients with pneumoconiosis were selected as case. The control group was workers exposed to dust but without pneumoconiosis who had the same sex, nationality, and workshop or work site as case. The differences in the age and cumulative exposure time between the case and control group were not move than five years and two years, respectively. The case matched with the control according to 1:1. Polymerase chains reaction-restriction fragment length polymorphism was used to determine the frequencies of TGF-beta genes in the two groups.

RESULTSThe frequencies of this TGF-beta (-509) genotypes were CC (22.2%), CT (43.6%) and TT (34.2%) in cases, which was significantly different from the control group, respectively (OR = 1.390, P < 0.05). There was no significant difference for frequency of TGF-beta+869 genotypes and allelic between case and control (P > 0.05). The frequencies of the TGF-beta (+915) genotypes in case [GG (70.9%), GC (29.1%)] were significantly different from the control group (OR = 1.455, P < 0.05). The frequency of TGF-beta (+915) * C allele in the case and control was 14.5% and 8.5%, respectively (P < 0.05). The frequencies of carrying TGF-beta (-509) CC and (+915) GG genotypes were 12.8% and 29.9% in case and control. The frequencies of carrying TGF-beta (-509) * T and (+915) * C alleles were 9.8% and 5.1% in pneumoconiosis and control (P < 0.05).

CONCLUSIONSTGF-beta (-509)CC genotype may be the protective factor for the pneumoconiosis. TGF-beta (+915)GC genotype may be a susceptible factor for the pneumoconiosis. The workers of carrying TGF-beta (-509) * T and (+915) * C alleles are more susceptible to pneumoconiosis.

Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Pneumoconiosis ; genetics ; Polymorphism, Single Nucleotide ; Transforming Growth Factor beta1 ; genetics

OBJECTIVETo establish a method for gene delivery in murine renal tissue using lentivirus vector encoding miR-483-5p.

METHODSThirty-five C57BL/6J mice were randomly divided into control group, low-dose treatment group (5 µL each kidney) , and high?dose treatment group (20 µL each kidney), and in the latter two groups, the lentivirus vector encoding miR-483-5p were injected in the renal cortex. The tissue samples were collected at 7 and 21 days after the injection. A transgenic mouse model with inducible systemic overexpression of miR-483-5p was established in TG483 mice. The Cre-loxp system was used to create a mouse model with renal tubule-specific expression of miR-483-5p. The levels of BUN in the mice were detected and HE staining and fluorometric TUNEL assay were used to observe the morphological changes of the kidneys; real-time qPCR was used to detect miR-483-5p expression in the renal cortex.

RESULTSThe mice with overexpression of miR-483-5p had normal renal function without obvious pathological changes or apoptosis in the renal tissue. Renal cortex injection of 20 µL lentivirus resulted in obviously increased level of miR-483-5p at 21 days (1.2∓0.43 vs 8.6∓1.09, P<0.001). miR-483-5p showed a low expression (0.9∓0.09 vs 1.7∓0.19, P<0.05) in TG483 mice and a high expression in the kidney of the transgenic mice established using the Cre-loxp system (1.6∓1.13 vs 12.36∓3.89, P<0.05).

CONCLUSIONThe transgenic mice with renal tubule-specific expression of miR-483-5p show normal renal function, and this model facilitates further study of the role of miR-483-5p in the kidney.

OBJECTIVETo investigate the effect of estradiol on the expression of antioxidant enzymes in osteoblasts and its role in postmenopausal osteoporosis.

METHODSRat models of osteoporosis established by ovariectomy were treated with estradiol for 3 months, and the changes in serum levels of reactive oxygen species (HO) and antioxidant enzymes (γ -GCS, GSH-ST and GSH-px) were detected. The effects of estradiol on the expression of γ -GCS mRNA and protein in osteoblast-like cells MC3T3-E1, MG63 and OB were examined with PCR and Western blotting. Using a mRNA microarray, we analyzed the changes in the expressions of 84 antioxidant enzymes in the osteoblast cell line MC3T3-E1 following estradiol treatment, and the enzymes with significant changes were verified by PCR. CCK-8 kit was used to evaluate the effect of estradiol and antioxidant NAC on the proliferation of MC3T3-E1 cells.

RESULTSRat models of osteoporosis were successfully established with ovariectomy. The osteoporotic rats showed significantly increased serum level of reactive oxygen species (H2O2) and decreased levels of antioxidant enzymes. Estrogen treatment of the osteoporotic rats obviously reversed the phenotype of osteoporosis, lowered serum level of reactive oxygen species, and increased the level of γ -GCS. In MC3T3-E1, MG63 and OB cells, estradiol treatment significantly upregulated the expression levels of γ -GCS mRNA and protein. In MC3T3-E1 cells treated with estrogen, the mRNA chip identified 6 upregulated antioxidant enzymes (Gpx6, Gstk1, Nos2, Prdx2, Ngb and Ccs), and the results of PCR verified that estradiol upregulated Ccs and Ngb mRNAs in MC3T3-E1, MG63 and OB cells. Estradiol and antioxidant NAC obviously promoted the proliferation of MC3T3-E1 cells.

CONCLUSIONEstradiol significantly increases the expression of antioxidase γ -Gcs, Ccs and Ngb in osteoblasts in vitro. Postmenopausal osteoporosis is closely related with the increase of reactive oxygen species and the decrease of antioxidant levels. In osteoblasts, estrogen deficiency may increase the level of reactive oxygen species, decrease the level of antioxidant enzymes, activate the oxidative stress cascade, and consequently inhibit the proliferation of osteoblasts to aggravate the condition of osteoporosis.

Objective To find out about the sanitary conditions of drinking water along the border line of Inner Mongolia via safety evaluation of drinking water along the border line of Inner Mongolia.Methods 108 samples of drinking water along the border line of Inner Mongolia(70 samples of self-supply source water,21 ones of self-supply tap water and 17 ones of municipal tap water)were collected.Hygienic evaluation of sensory indexes, normal chemical indexes, toxicological indexes and microbiology indexes of water quality was performed according to hygienic standards for drinking water(GB 5749—2006).Results 58 samples were qualified with a total qualified rate of 53.7%.The unqualified rate of the total number of coliforms was the highest(20.4%),followed by fluoride(19.4%).As for deep wells, shallow wells and surface water,there was no statistically significant difference.Compared with municipal tap water,the unqualified rate of self-supply water(28.6%)was higher(17.7%).Conclusion The qualified rate along the border line of Inner Mongolia of drinking water is low.Treatment and disinfection facilities for drinking water,detection devices of water quality are needed.The cleaning and disinfection of storage tanks should be performed periodically in oder to prevent waterborne infectious diseases.

Objective To investigate children with nephrotic syndrome (NS) receiving long-term glucocorticoid treatment while monitoring insulin resistance(IR) index,and analyze the influencing factors,and to observe the efficacy of metformin for preventing glucocorticoid-induced obesity and diabetes mellitus type 2.Methods A total of 32 cases including 20 boys and 12 girls with mean age of 7.6 years old(2 to 14 years old),were diagnosed as NS.All children undergoing long-term treatment of prednisone ＞ 1 mg/(kg · d) for at least 3 months.The height,body weight and body mass index(BMI) were monitored.The concentrations of serum insulin and C-peptide were determined by radioimmunoassay,the concentrations of serum HbA1 c were determined by high performance liquid chromatography,and the levels of fasting glucose,cholesterol and triglyceride were measured with an automatic measuring analyzer (American ABBO TT CCX-Ⅱ).The homeostasis model assessment-estimated insulin resistance(HOMA-IR) was calculated;When HOMA-IR ＞ 3.5,the patients were given metformin treatment for 12 weeks while supervising their height,weight,fasting blood glucose levels monthly.Results Out of 32 cases of NS with long-term use of prednisone,23 cases had HOMAIR ＞ 3.5 (72％).Thus,IR group included 23 patients,and non-IR (NIR) group had 9 patients.There were significant differences between 2 groups in BMI,age,prednisone dosage and the levels of triglyceride (TG) (all P ＜ 0.05).After 12 weeks of metformin treatment,the HOMA-IR (5.24 ± 1.82 vs 2.54 ± 1.09,P ＜ 0.01),HbA1 c levels [(6.36 ±0.82)％ vs(5.39 ±0.51)％,P ＜0.05],BMI[(27.42 ±6.12) kg/m2 vs(22.72 ±5.48) kg/m2,P ＜0.01],and TG levels[(2.03 ± 1.10) mmol/L vs(1.45 ±0.48) mmol/L,P＜0.05] were significantly reduced.Conclusions Older children with long-term use of glucocorticoid may easily lead to IR.Therefore,monitoring the HOMA-IR and early administration of mefformin is helpful for prevention and treatment of glucocorticoid-induced obesity and diabetes mellitus type 2.