BackgroundRetinal pigment epithelial(RPE) cells can secrete platelet-derived growth factor (PDGF) and PDGF receptor(PDGFR).Studies have shown that PDGF plays a key role in the formation of proliferative vitreous retinopathy(PVR). ObjectiveThis study was to investigate the proliferation and apoptosis changes of RPE after blockage of the PDGFR-α expression by antisense oligonucleotide ( ASODN ) in vitro. Methods Human RPE cells strain was cultured in low glucose DMEM with 10％ fetal bovine serum.Logarithmic phase cells were collected and incubated in 96-well plate at the density of 5 × 105 cells/hole.PDGFR-α ASODN was transfected into RPE cells at different concentrations for 48 hours.The cells of the blank control group were regularly cultured without any transfection.The changes of PDGFR-α expression were detected by reverse transcription-polymerase chain reaction(RT-PCR),and the proliferation of RPE was detected by MTT as the A490 value.Hoechst 33258 fluorescence staining was used to determine the apoptosis of RPE.Flow cytometry method (FCM) was applied to detect the change of cell cycle and apoptosis rate of RPE cells. ResultsThe A490 values of RPE cells were 1.45±0.12,1.07±0.06,0.65±0.05 in blank control group,1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group with the significant difference(P=0.00 ),and that of 1.0 μmol/L Lipo-ASODN group and 2.0 μ mol/L Lipo-ASODN group were significantly lower than the blank control group ( P =0.00,0.00).Hoechst 33258 staining showed that the apoptosis cells were obviously more in Lipo-ASODN group compared with blank control group.PDGFR-α ASODN transfection induced an increase of percentage of RPE cells in G0/G1 phase( F =206.70,P =0.00),and the apoptosis rates in 1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group were significantly enhanced in comparison with blank control group ( 37.8 ± 1.3 vs 10.5 ± 0.1,61.2 ± 1.9 vs 10.5 ± 0.1 ) ( F =1808.90,P =0.00 ).Expression intensity of PDGFR-α mRNA in RPE cells in Lipo-ASODN groups was lower. ConclusionsBlocking the PDGFR-α expression with ASODN technology can suppress proliferation and induce apoptosis of RPE cells.Intensity of PDGFR-α mRNA expression in RPE cells is ASODN dose-dependent.ASODN targeted to PDGFR-α offers an experimental basis of the gene therapy for PVR.

Objective To explore the endoplasmic reticulum stress (ERS) apoptosis pathway in unilateral ureteral obstruction (UUO) mechanism of renal interstitial fibrosis in rats.Methods The rats were randomly divided into con-trol group and model group,UUO group line ligation of the left ureter,three days 3,7 and 14 days HE and Masson staining were used to observe the renal pathological changes;Take blood retinal venous plexus,separat and determina-tion of serum blood urea nitrogen and creatinine;Western blot was used to find protein 78 (GRP78) glucose regula-tion,endoplasmic reticulum source sex transcription factor (CHOP) and apoptosis related proteins cysteine aspartic acid proteinase 3(caspase-3) and caspase-12 protein expression. Results Compared with control group,the visible UUO model group 1) expansion of renal tubule and renal interstitial fibrosis degree with the extension of ureteral ob-struction time and progressive increase;2) GRP78,CHOP,caspase-3 and caspase-12 protein expression in postoper-ative 3 d have to rise,as the obstruction prolonged,the protein expression more significantly(P<0.01).Conclusions Endoplasmic reticulum stress related trademark protein in UUO rat renal interstitial fibrosis is the increase in expres-sion may promote the early renal interstitial fibrosis and continuous progress.

Objective To explore the endoplasmic reticulum stress (ERS) apoptosis pathway in unilateral ureteral obstruction (UUO) mechanism of renal interstitial fibrosis in rats.Methods The rats were randomly divided into con-trol group and model group,UUO group line ligation of the left ureter,three days 3,7 and 14 days HE and Masson staining were used to observe the renal pathological changes;Take blood retinal venous plexus,separat and determina-tion of serum blood urea nitrogen and creatinine;Western blot was used to find protein 78 (GRP78) glucose regula-tion,endoplasmic reticulum source sex transcription factor (CHOP) and apoptosis related proteins cysteine aspartic acid proteinase 3(caspase-3) and caspase-12 protein expression. Results Compared with control group,the visible UUO model group 1) expansion of renal tubule and renal interstitial fibrosis degree with the extension of ureteral ob-struction time and progressive increase;2) GRP78,CHOP,caspase-3 and caspase-12 protein expression in postoper-ative 3 d have to rise,as the obstruction prolonged,the protein expression more significantly(P<0.01).Conclusions Endoplasmic reticulum stress related trademark protein in UUO rat renal interstitial fibrosis is the increase in expres-sion may promote the early renal interstitial fibrosis and continuous progress.

Objective To investigate the therapeutic methods of hyperpotassemia induced by excessively high blood concentration of tacrolimus (FK506) caused by drug use after renal transplantation. Methods Clinical data of 10 patients diagnosed with hyperpotassemia induced by excessively high blood concentration of FK506 after administration of antifunga l medication following renal transplantation were collected and retrospectively analyzed. Results At 1-2 months after renal transplantation, 10 patients suffered from pulmonary infectiono r pneumonia complicated with pulmonary fungal infection . An appropriate dose of compound sulfamethoxazole, micafungin, cefoperazone sodium-sulbactam sodium and moxifloxacin was administered for antifungal infection. After potassium-lowering therapy, termination of antifungal medication and FK506 dose adjustment (replaced by cyclosporin for certain cases), the serum level of potassium was declined and maintained within normal range for 10 cases. The serum concentration of FK506 was within normal range. No sign of excessively high level of potassium was observed without any potassium-lowering intervention. Conclusions Postoperative administration of drugs is likely to cause excessively high level of FK506 and hyperpotasesmia. Potassium-lowering therapy, termination of drug use and adjustment of immunosuppressive agents should be adopted to avoid the incidence of adverse pharmacologic interaction.

Objective To investigate the effectiveness and safety of rosuvastatin and simvastatin in the treatment of hyperlipidemia. Methods The database including CNKI, VIP, Wanfang data base and CBM were retrieved to search the clinical randomized controlled trials (RCT) of rosuvastatin and simvastatin in the treatment of hyperlipidemia, and the data were analyzed with Review Manager 5.2. Results Eighteen RCTs were included with a total sample size of 1819 cases with hyperlipidemia, in which there were 917 patients in rosuvastatin group and 902 in simvastatin group. The Meta-analysis results showed that there were significantly lower serum levels of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), triglyceride (TG) and significantly higher level of high-density lipoprotein cholesterol (HDL-C) in rosuvastatin group compared with those of simvastatin group [(MD=-0.15, 95%CI:-0.22--0.09, P<0.01), (MD=-0.18, 95%CI:-0.25--0.11, P<0.01), (MD=-0.23, 95%CI:-0.28--0.19, P<0.01) and (MD=-0.11, 95%CI:-0.06--0.15, P<0.01)]. There was no significant difference in the incidence of gastrointestinal adverse reaction between the two groups. Conclusion The current clinical evidences show that rosuvastatin has a better effect on the treatment of hyperlipidemia, and has no adverse reactions.

OBJECTIVETo investigate the association of bone metabolism related genes polymorphisms with the effect of raloxifene hydrochloride(RLX) on bone mineral density (BMD) and bone turnover markers in postmenopausal women with osteoporosis.

METHODSA total of 68 unrelated postmenopausal women with osteoporosis of Han ethnicity aged 47-74 years were randomly divided into 2 groups of 34 women: RLX group (60 mg were given daily for 12 months) and placebo group. BMD and bone turnover markers were measured at baseline, 6 and 12 months after treatment. The polymorphisms of Xba I and Pvu II sites in estrogen receptor 1 gene(ESR1), Ras I site in ESR2 gene, and start codon (Fok I) and CDX2 binding sites in vitamin D receptor gene (VDR) were analyzed.

RESULTSA total of 58 patients completed 12 months of study period. By the end of study, the increased percentage of BMD in lumbar spine 2-4 (L2-4), total hip, and trochanter were found significantly different between RLX group and placebo group(P<0.05), and the decreased percentage of C-telopeptide and osteocalcin were significantly different between the two groups (P<0.01). The BMD of total hip and trochanter of women with FF genotypes of VDR Fok I site were decreased by 1.98%+/-4.86% and 2.26%+/-4.73% respectively in the RLX group, but those of women with Ff/ff genotypes were increased by 2.52%+/-2.75% and 2.74 %+/-2.97%, respectively(P<0.05). Moreover, the total hip BMD of women with PP/Pp genotypes of ESR1 Pvu II site was increased by 2.12%+/-2.78%, and of women with pp genotype it was decreased by 1.34%+/-3.73%(P<0.05). However, no significant association was observed of the polymorphisms of five sites with the changes of BMD and bone turnover markers in the placebo group.

CONCLUSIONThe effect of RLX on BMD in postmenopausal women with osteoporosis is regulated by the polymorphisms of Fok I of VDR gene and Pvu II of ESR1 gene. The study is valuable to select this drug according to genotype of patients in clinical.

Aged ; Biomarkers ; metabolism ; Bone Density ; drug effects ; genetics ; Bone Diseases, Metabolic ; genetics ; metabolism ; Bone Remodeling ; drug effects ; genetics ; Bone and Bones ; drug effects ; Double-Blind Method ; Female ; Humans ; Middle Aged ; Osteoporosis ; drug therapy ; Osteoporosis, Postmenopausal ; drug therapy ; Polymorphism, Genetic ; Postmenopause ; drug effects ; Raloxifene Hydrochloride ; pharmacology ; therapeutic use ; Selective Estrogen Receptor Modulators ; pharmacology ; Women

This study was aimed to explore the prognostic significance of telomere length in patients with chronic lymphocytic leukemia (CLL) and to analyze relation of telomere length with Binet stage, IgVH mutation status, CD38, ZAP-70 expression as well as other clinical features. 35 CLL patients who contained 80% or more tumor cells in the peripheral blood or bone marrow samples were selected as objects studied, while 13 healthy donors were served as normal controls. The telomere relative length was detected by using a real-time fluorescent quantitative polymerase chain reaction method (qPCR); the expression of CD38 and ZAP-70 protein were detected by flow cytometry, the IgVH mutation was detected by multiplex PCR. The results showed that the mean telomere relative length in CLL patients and normal controls were 0.384 and 0.443 respectively, but the difference between them was not significant (p > 0.05). The telomere length was significantly correlated with Binet stages and IgVH mutation status. Patients in Binet stage B and C showed significantly shorter telomeres than those in Binet stage A (p = 0.001). Mean telomere relative lengths in patients without IgVH mutation were shorter than those in patients with IgVH mutation (p = 0.015). No relation of telomere length with sex, age, ZAP-70 protein and CD38 were found (p > 0.05). It is concluded that telomere length may have a prognostic significance for CLL patients. Combining telomere length and IgVH mutation status may achieve a better prognostic subclassification for CLL patients.
ADP-ribosyl Cyclase 1 ; metabolism ; Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Female ; Humans ; Leukemia, Lymphocytic, Chronic, B-Cell ; genetics ; metabolism ; Male ; Middle Aged ; Mutation ; Prognosis ; Telomere ; chemistry ; genetics ; metabolism ; ZAP-70 Protein-Tyrosine Kinase ; metabolism

OBJECTIVETo investigate the association of Apa I polymorphism in vitamin D receptor (VDR) gene with bone mass in men.

METHODSThe VDR Apa I genotype was determined by PCR-restriction fragment length polymorphism (RFLP) in 388 unrelated healthy men aged 46-80 years of Han nationality in Shanghai city. Bone mineral density (BMD) and bone mineral content (BMC) at lumber spine 1-4 (L1-4) and at any sites of proximal femur including to femoral neck (Neck), trochanter (Troch) and Ward's striangle (Ward's) were measured by duel-energy X-ray absorptiometry.

RESULTSFrequencies distribution of VDR Apa I genotype were aa for 48.1%, Aa for 44.2% and AA 7.7%. The allele frequencies of Apa I polymorphism were in Hardy-Weinberg equilibrium. No significant association was found between Apa I genotype and BMD or BMC in group of all population or in subgroup of men below 60 years. In men above 60 years, the significant association was found between VDR Apa I genotype and BMD or BMC at L1-4, Neck and Ward's (P < 0.05, P < 0.01) and compared with Aa and aa genotype, AA genotype had significantly higher mean BMD and BMC at L1-4, Neck and Ward's (P < 0.05, P < 0.01). But Apa I genotype is not associated with BMD and BMC at Troch.

CONCLUSIONSApa I polymorphism is associated with bone mass in men above 60 years, and AA genotype has higher bone mass. Apa I polymorphism in VDR gene possibly influence loss of trabecular and cortical bone mass in old men.

Age Factors ; Aged ; Aged, 80 and over ; Alleles ; Bone Density ; Humans ; Male ; Middle Aged ; Phenotype ; Polymorphism, Restriction Fragment Length ; Receptors, Calcitriol ; genetics ; Transcription Factors ; genetics ; Zinc Fingers ; genetics

OBJECTIVETo investigate the relationship of Msp AI polymorphism in the promoter region of cytochrome P450c 17alpha (CYP17) gene with bone mass and bone size in Shanghai men of Han nationality.

METHODSThe CYP17 Msp AI genotype was determined by polymerase chain reaction-restriction fragment length polymorphism in 397 unrelated men (324 healthy men, 73 osteoporosis patients) aged 46-80 years of Han nationality in Shanghai. Bone mineral density (BMD), bone mineral content (BMC), and bone cross-section area (CSA) at lumber spine 1-4 and at any sites of proximal femur, including femoral neck, trochanter and Ward's triangle were measured by duel-energy X-ray absorptiometry.

RESULTSFrequency distributions of CYP17 genotype were TC (51.1%), CC (33.8%), and TT (15.1%). The allele frequencies T and C were 40.7% and 59.3%, respectively. Allele frequencies did not deviate from Hardy-Weinberg equilibrium. The frequencies of CYP17 Msp AI genotype did not show difference between osteoporosis cases and healthy controls. In group of all population, or in subgroups of osteoporosis patients and healthy men, CYP17 Msp AI genotype was not significantly associated with BMD, BMC, and CSA at lumber spine 1-4 and at any sites of proximal femur after having been adjusted for age, weight, and height with analysis of covariance.

CONCLUSIONMsp AI polymorphism of CYP17 gene is not a genetic factor that influence the variation of bone mass and bone size in Shanghai men of Han nationality.

Aged ; Aged, 80 and over ; Alleles ; Asian Continental Ancestry Group ; Bone Density ; China ; Femur ; anatomy & histology ; pathology ; Gene Frequency ; Humans ; Lumbar Vertebrae ; anatomy & histology ; pathology ; Male ; Middle Aged ; Osteoporosis ; genetics ; pathology ; Phenotype ; Polymorphism, Restriction Fragment Length ; Promoter Regions, Genetic ; genetics ; Steroid 17-alpha-Hydroxylase ; genetics

OBJECTIVETo investigate the role and significance of FHIT genes depletion, p53 overexpression and HPV16/18 infection in cervical intraepithelial neoplasia (CIN) and cervical carcinoma (CC).

METHODSTumor samples taken from 52 cases of CIN and 69 cases of CC were processed by immunohistochemistry (SP) to determine the expression of FHIT genes and p53 protein, by in situ hybridization to detect HPV16/18 infection, and were compared with those in 18 cases of normal cervical tissues as control.

RESULTS(1) The FHIT expression was positive in normal cervical tissue with no depletion occurred, and was 30.8% in CIN. It was significantly higher in CIN III and carcinoma groups than that in normal and CIN I/II groups (P < 0.01). The depleted expression of FHIT in infiltrating cervical carcinoma group was 66.7% (46/69), significantly higher than that in normal and CIN groups (P < 0.01). Along with the decreasing of cell differentiation, the negative rate of FHIT raised. (2) The positive expression of p53 in CC group was 56.5% (39/69) and the HPV16/18 was 84.1% (58/69), both higher than that in CIN and normal groups (P < 0.05). (3) In CIN and CC groups, the positive rate of p53 in cases with positive or negative FHIT expression was similar (P > 0.05). (4) There is a negative correlation between FHIT and p53 expression. The rate of HPV16/18 infection in the depleted expression of FHIT group was significantly higher than that in FIHT normal expression group (P < 0.01).

CONCLUSION(1) The FHIT-depletion is related with cervical carcinogenesis. It may be used as a marker to serve mass screening of CIN-high risk subjects and diagnostic indicator for early cervical carcinoma. (2) Depleted expression of FHIT is frequently associated with p53 over-expression in CIN and CC subjects, but there is no direct correlation between them. (3) HPV16/18 infection may probably be the common cause leading to altered FHIT and p53 expression.

Acid Anhydride Hydrolases ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; virology ; Cervical Intraepithelial Neoplasia ; metabolism ; virology ; Female ; Human papillomavirus 16 ; genetics ; Human papillomavirus 18 ; genetics ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Neoplasm Proteins ; metabolism ; Papillomavirus Infections ; metabolism ; virology ; Tumor Suppressor Protein p53 ; metabolism ; Uterine Cervical Neoplasms ; metabolism ; virology