Biological Clocks ; Genetic Therapy ; Heart Diseases ; therapy ; Humans ; Stem Cell Transplantation

Background Congenital cataract is an important cause of blindness and amblyopia in children,and about 50％ of congenital cataract is hereditary.Objective The aim of this study was to determine the diseasecausing gene of one Hui congenital cataract pedigree by using exon combined target region capture sequencing chip of eye diseases.Methods This study was approved by Ethic Committee of Ningxia People's Hospital and followed Declaration of Helsinki.One Hui congenital cataract pedigree was recruited in Ningxia Eye Hospital in 2011.All the disease history of the members in this family were collected and recorded,and the eye examinations were performed.The peripheral blood specimens were collected from family members and 300 healthy individuals for the extraction of DNA.Exon combined target region capture sequencing chip of eye diseases was used to screen the candidate diseasecausing mutations,then PCR and direct sequencing were used to confirm the disease-causing mutations.Results This H ui family included 61 members of 6 generations,and 18 patients were diagnosed in serial 5 passages,conforming to autosomal dominant inheritance pattern.Among 18 cataract patients,7 individuals were associated with nystagmus and strabismus,and 4 patients had high myopia.Eight candidate pathogenetic mutations were detected by exon combined target region capture sequencing chip of eye diseases and bioinformatics method,with 5 mutations in noncoding regions and 3 in coding regions.The mutation P24T of CYRGD gene was confirmed as pathogenic mutation of this pedigree by using PCR and direct sequencing methods.These mutations co-segregated with affected members of the family,and the mutations were not found in the unaffected family members and 300 unrelated controls.Conclusions P24T of CYRGD gene mutation is confirmed as pathogenic mutation of this pedigree.Exon combined target region capture sequencing chip provides a new approach to detect disease-causing mutations of congenital cataract with diversity clinical phenotypes.

BACKGROUND:Some scholars have found that cervical vertebral body bone trabecula was reduced,became thin,even perforated in old patients with osteoporosis.Whether this change will induce cervical vertebral body deformation,and what relationship to the onset of cervical syndromeOBJECTIVE:To study the relation of cervical spondylotic myelopathy and osteoporosis by measuring and comparing.METHODS:A totaI of 40 subjects with normal lumbar vertebra density and without cervical spondylosis were enrolled as control group,averagely 32 5 years.A total of 30 patients with cervical spondylosis served as cervical spondylosis group,averagely 43.6 years.Totally 46 patients with cervical spondylosis and osteoporosis served as combined with osteoporosis group,averagely 58.6 years.116 subjects underwent radiograph Height and sagittal diameter of the vertebral body ratio of height to sagittal diameter of the vertebral body.and ratio of sagittal diameter of cervical canal/vertebra body were measured.RESULTS AND CONCLUSION:Compared with the control group.vertebral height was decreased.and sagittal diameter became longer(P＜0.05),and the ratio of sagittal diameter of cervical canal/vertebra body became smaller(P＜0.05)in the combined with osteoporosis group.Vertebral body deformation was characterized by decreased vertebral height and prolonged sagittal diameter became flat.Results suggested that osteoporosis induced cervical vertebral deformation,correlation between osteoporosis and cervical spondylosis,which may be a factor for cervical spondylosis development.

Objective: To denvolop HPLC an method for the determination of the content of three saponins in Zhixiao suppositories. Methods: Analysis carriedout was performed on a Kromasil C18 column (4.6 mm×250 mm., 5μm)at 23℃. The mobile phases were methanol and 0.2% phosphoricacid solution in a gradient elution. The flow rate was set at 1.0 mL·min-1 and the ultraviolet detector was operated at 203 nm. Results: The linear ranges of notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1 were 0.368～2.76μg,0.676～5.07 μg and 0.812～6.09μg,respectively. The average recoveries of the three saponins were 98.53%, 99.20% and 99.35%, respectively. The RSD (n=5) were 2.11%, 1.88% and 2.38%, respectively. Conclusion: The separation is a good and the results are accurate and reproducible. It can be used for the quality control of Zhixiao suppositories.

Objective To investigate the effect and safety of sildenafil on persistent pulmonary hypertension of the newborn (PPHN). Also compared the effect of sildenafil with tolazoline and milrione. Methods Forty five neonates with PPHN were recruited from January 2005 through October 2008 in NICU, 25 males and 20 females. The median gestational age was (39.3 + 2.4) weeks, the median birthweight was (3 114.0±10.2) g, and the median age were (13.0±0.8) hours. The patients were randomly assigned to receive sildenafil, tolazoline and milrione therapy. The pulmonary artery pressure (PAP) was measured by echocardiography. Results Thirty patients were cured, 6 patients were improved and 9 patients were of no effect. The total effective rate was 80%. There was no statistical difference among sildenafil, tolazoline and milrione. The PAP decreased when the patients were treated with sildenafil, tolazoline and milrione. No side effects happened in all patients treated with the three drugs. Conclusions Sildenafil is an effective and safe drug to reduce PAP of PPHN and it also help to improve cardiac function.

ObjectiveTo observe the subcutaneous and intraperitoneal insulin injection's effect of the level of oxygen free radicals of type 2 diabetes model.MethodsC57BL/6J mice were chosen as normal control group (C group,n=9).KK mice were randomly divided into intraperitoneal injection of insulin group (i.p.group,n=9),the subcutaneous insulin group (s.c.group,n=9) and untreated group (U group,n =9).The i.p.group and the s.c.groups were given certain amount of insulin (insulin injecta and protamine insulin injecta by volume ratio of 2:1 mixture)for one month,maintained the GLU at normal levels (6±1.5) mmol/L.SOD,GSH-PX activity and MDA content of serum,liver,kidney and heart in each group were detected.Results The liver,kidney,heart and serum's SOD and GSH-PX activity significantly reduced and MDA content significantly increased in the U group.Both kinds of delivery methods could increase serum SOD and GSH-PX activity and reduce the content of MDA to the normal control group level,but the intraperitoneal injection had stronger effect.Two kinds of delivery methods could both reduce the MDA content of liver,and had almost the same effect; but the subcutaneous injection group had better effect on increasing the liver's SOD activity,and the intraperitoneal injection had better effect on increasing liver's GSH-PX activity.Intraperitoneal injection had better effect on reducing kidney' s MDA content and increased SOD activity.Two kinds of delivery methods had the same effect on reducing the heart's MDA content.Conclusion The two delivery methods can both make the MDA levels of KK mice in serum,heart,liver and kidney fall to as normal as that of control group,but the two delivery methods have different ways of improving the antioxidant capacity in different organs.Intraperitoneal injection can reduce MDA content in serum and kidney better.

OBJECTIVETo investigate the changes of CD4 + CD25 + Foxp3 + regulatory T cells in the peripheral blood mononuclear cells (PBMC) and their association with insulin resistance in different stages of prostate cancer (PCa).

METHODSUsing flow cytometry, we counted the CD4+ CD25 + Foxp3 + regulatory T cells in the PBMCs of 62 PCa patients (5 cases of TNM stage I, 16 cases of stage II, 21 cases of stage III, and 20 cases of stage IV) and 42 normal healthy controls, and calculated their proportion in the CD4+ T-lymphocytes. We determined the levels of fast blood glucose (FBG) and fast insulin (FINS) for the insulin resistance index (HOMA-IR), obtained the serum IGF-1 level by ELISA, and analyzed the relationship of the count and proportion of CD4+ CD25+ Foxp3+ regulatory T cells with insulin resistance by comparison between the PCa patients and normal healthy controls.

RESULTSCompared with the control group, the PCa patients showed significantly increased HOMA-IR (3.68 ± 1.42 vs 6.68 ± 1.66), decreased level of serum IGF-1 ([164.56 ± 30.58] vs [96.39 ± 21.21] ng/ml), and elevated count ([1.99 ± 0.78 ] x 10(7) vs [3.55 ± 0.29] x 10(7)) and proportion ([5.33 ± 0.65] vs [13.88 ± 0.96]%) of CD4 + CD25 + Foxp3 regulatory T cells in the PBMCs. The TNM stage was correlated positively with the count and percentage of CD4 + CD25+ Foxp3 + regulatory T cells and HOMA-IR, but negatively with the level of serum IGF-1. Meanwhile, the count and percentage of CD4 + CD25 + Foxp3 + regulatory T cells were found to have a positive correlation with HOMA-IR (r = 0.722 and 0.689, P < 0.01) but a negative correlation with the level of serum IGF-1 (r = -0.747 and -0.896, P < 0.01).

CONCLUSIONThe count and proportion of CD4+ CD25 + Foxp3 + regulatory T cells in the peripheral blood and insulin resistance increase with the elevated stage of PCa. CD4 + CD25 + Foxp3 + regulatory T cells may be involved in the occurrence and progression of PCa by regulating insulin resistance.

Aged ; CD8-Positive T-Lymphocytes ; Case-Control Studies ; Disease Progression ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Humans ; Hyperinsulinism ; Insulin ; blood ; Insulin Resistance ; Insulin-Like Growth Factor I ; metabolism ; Leukocytes, Mononuclear ; Lymphocyte Count ; Male ; Prostatic Neoplasms ; blood ; immunology ; pathology ; T-Lymphocytes, Regulatory

Objective To study the expression of anticentromere antibody (ACA)on multiple diseases to help the clinical diag-nosis and treatment.Methods The clinical and laboratory data of 129 cases with positive ACA were retrospectively ana-lyzed.Results ①116 females and 13 males were found among 129 cases with positive ACA,and serology titer of ACA in fe-male patients was higher than male and more compound antibody were shown in female patients,which was significantly dif-ferent compared with males (χ2=6.28,P=0.01;χ2=6.85,P=0.003).②Among 118 cases which had detailed clinical in-formation,there were 65 patients suffered from autoimmune diseases (55.08%)and 53 cases suffered from non-autoimmune diseases (44.92%).Autoimmune disease cases showed more compound and high serology titer,there were significant differ-ence between the two groups (χ2=21.97,P<0.001;χ2=11.44,P=0.001).Conclusion Though ACA was rarely found,it was shown in autoimmune and non-autoimmune diseases and can be not too severe or fetal and even lead to multi-organ fail-ure.So it should be taken seriously.

Objective: To observe the changes of liver metabolism in mice exposed to artificial light at night. Methods: Healthy male C57BL/6J mice were randomly divided into the light at night group and the control group, with 8 mice in each group. The daily light/dark cycle was 12/12 hours in the control group, and 24/0 hours in the light at night group for 10 consecutive days. The hepatic metabolite profiles of the two groups of mice were detected by high performance liquid chromatography tandem mass spectrometry. The modelling was assessed by combining principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis ( OPLS-DA ) , The changes of metabolites in the two groups were compared through KEGG database. Results: Compared with the control group, 9 different metabolites were detected in the light at night group, among which the down-regulated metabolites were glycine-betaine, glutathione, tyrosine, betaine, lysine, hypoxanthine, histidine and methionine, and the up-regulated ones were mannose-6-phosphate. The weight analysis of the metabolic pathways showed that the major influences on liver of light at night group were phenylalanine-tyrosine-tryptophan metabolism, tyrosine metabolism, fructose and mannose metabolism, cysteine and methionine metabolism and histidine metabolism. Conclusion The metabolism of various amino acids and sugars in light at night mice is disturbed,and the key differential metabolites are tyrosine, methionine, histidine and mannose-6-phosphate.

Objective To determine the effect of deferoxamine administration on autophagy in a rat model of blast-induced brain injury.Methods Thirty-nine male SD rats were allotted to shamoperated group,injury group and deferoxamine group with 13 rats in each,according to the random number table.Feeney's method was applied to establish the model.Deferoxamine group received deferoxamine of 100 mg/kg intraperitoneally.Sham-operated and injury group were injected with saline intraperitoneally.All treatments were started two hours postinjury at 12 hour interval for up to 28 days.Hemoglobin,rectal temperature,blood glucose and mortality were detected at 1,3,7,14 and 28 days.Morris water maze was conducted.Rats were killed later for detecting the brain defect volume and level of Beclin 1 at the site of injury.Results There were no significant differences among the three groups with respect to hemoglobin,rectal temperature and blood glucose (P ＞ 0.05).Mortality in injury versus deferoxamine groups did not differ significantly (P ＞ 0.05).Volume of defected brain in deferoxamine group was (115.35 ± 13.70) mm3,smaller than (209.99 ± 16.70) mm3 in injury group (P ＜ 0.05).In Morris water maze test,the time spent in the searching the platform and latency to reach the platform were improved in deferoxamine group compared to those in injury group [(3.13 ± 0.35) vs (2.13 ± 0.64);(36.15 ± 26.63) s vs (110 ± 47.34) s respectively] (P ＜ 0.05).Both immunohistochemisty and western blot showed dramatically increased level of Beclin 1 after injury,but treatment with deferoxamine significantly reduced the Beclin 1 expression.Conclusion Level of Beclin 1 is significantly upregulated after blast-induced brain injury in rats,resulting in elevated autophagy postinjury,but the treatment with deferoxamine is neuroprotective possible by lessening autophagy damage.