Fibrosis ; Humans ; Inflammation ; MicroRNAs ; Neoplasms

Objective To investigate the early diagnostic value of cornel confocal microscopy for the screening of small neuropathy in elderly patients with type 2 diabetic mellitus.Methods In the prospective study,96 elderly patients with diabetes as study group and 46 patients with non-diabetes as the control group were continuously collected from our hospital endocrinology and ophthalmology out patients during May 2014 to February 2016.The 96 cases of type 2 diabetes were subdivided into 47 patients with diabetic peripheral neuropathy (DPN)and 47 patients with nowdiabetic peripheral neuropathy(non DPN).Results The diabetes course was shorter in non-DPN group than in DPN group(P=0.000).The levels of glycosylated hemoglobin and urine albumin were lower in the nonDPN than in the DPN(P =0.072,0.007,respectively).The corneal nerve fiber density was lower in the DPN group than in NDPN group (P =0.000).Corneal nerve fiber density was higher in control group than in DPN and NDPN group.The differences in number of corneal nerve fibers showed no statistical significance between DPN and NDPN group (x2 =2.391,P =0.314).But the number of corneal nerve fibers was significant less in DPN and NDPN group than in control group(x2 =16.014,P =0.000).The negative correlation was found between the course of disease and corneal fibrous density by using single factor linear regression analysis.The number of corneal nerve fibers was lower in smoking group than in non-smoking group(P=0.003).The multiple linear regression analysis showed that duration of diabetes was a risk factor for diabetic neuropathy.Conclusions In some elderly diabetic patients with non-neuropathy,corneal nerve fiber density and number have been significantly decreased before nerve conductive velocity is reduced.Therefore,corneal confocal microscopy can be used to detect and diagnose small diabetic neuropathy in elderly patients with diabetes mellitus.

Objective To compare the primary clinical results of the anteromedial portal (AMP) and accessory anteromedial portal (AAMP) techniques for femoral tunnel drilling in single-bundle anterior cruciate ligament (ACL) reconstruction.Methods Data of isolated ACL rupture patients who had undergone single-bundle ACL reconstruction with autologous semitendinosus and gracilis tendons from March of 2012 to February of 2014 were retrospectively analyzed.The femoral tunnels were drilled with AMP techniques in 14 patients (group AMP) and with AAMP techniques in 23 patients (group AAMP).All the patients were followed up for 6 to 29 months.At the latest follow-up the Lysholm,Tegner and international knee documentation committee (IKDC) scores were used to estimate knee joint function,while the Lachman test and Pivot-shift test were used to estimate knee joint instability.Results The average follow-up time was 16.07±7.31 months in group AMP and 13.35±5.92 months in group AAMP.In group AMP,the Lysholm,Tegner and IKDC average scores were 89.86±7.90,8.64±1.65 and 89.31±8.16,respectively.While they were 92.17±6.72,8.91±1.16 and 90.89±7.80 in group AAMP,respectively.In group AMP the Lachman test was negative in 11 patients and positive in 3 patients.In group AAMP the Lachman test was negative in 20 patients and positive in 3 patients.The Pivotshift test was negative in 9 patients,positive in 5 patients and negative in 20 patients,positive in 3 patients in group AMP and AAMP,respectively.There were no significant differences in Lysholm,Tegner,IKDC scores,the negative rates of Lachman and Pivot-shift tests between two groups.Conclusion Single-bundle ACL reconstructions using AMP and AAMP techniques for femoral tunnel drilling have similar excellent primary clinical results.

BACKGROUND: In this study, we have investigated the effect of Korean red ginseng (KRG) extracts on the production of TNF-alpha and IL-8 in human keratinocytes. Also, to examine the antioxidative effect of red ginseng extracts, free radical scavenging activity and superoxide dismutase (SOD) activity in human dermal fibroblasts was measured. METHODS: To investigate the effect of KRG in atopic dermatitis, we measured the level of TNF-alpha and IL-8 secretion in LPS-stimulated human keratinocytes after the treatment of KRG extracts using enzyme-linked immunosorbent assay. Anti-oxidative activity was investigated by measuring 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and SOD activity. RESULTS: The stimulation of human keratinocytes with KRG extracts shifted the LPS-induced cytokine secretion toward a more immunosuppressive response. KRG dose-dependently decreased TNF-alpha and IL-8 production in HaCaT cells and a significant inhibition of TNF-alpha was shown when cells were treated with 500 and 1,000 microg/ml of KRG extracts. Additionally, KRG extracts showed DPPH radical scavenging and SOD activity in a dose-dependent manner. Particularly, SOD activities of concentrations higher than 60 microg/ml of KRG extracts were significantly different in human dermal fibroblast cells. CONCLUSION: Based on this study, KRG extracts may be a useful immunosuppressive agent in the treatment of atopic dermatitis.
Biphenyl Compounds ; Dermatitis, Atopic ; Enzyme-Linked Immunosorbent Assay ; Fibroblasts ; Humans ; Interleukin-8 ; Keratinocytes ; Panax ; Picrates ; Superoxide Dismutase ; Tumor Necrosis Factor-alpha

Acanthopanax koreanum Nakai, a well known traditional herb grown in Jeju Island, South of Korea, has been used as a tonic and sedative agent, as well as in the treatment of diabetes and immune diseases. Mutagenicity of two lignans, syringaresinol and tortoside A isolated from A. koreanum, was assessed using Salmonella/microsome (Ames) test. Tester strains used were Salmonella typhimurium TA98, TA100, TA1535, and Escherichia coli WP2uvrA. The mutagenic activity was determined both in the absence or presence of S9 mixture. As a result, tortoside A did not cause any increase in the number of his+ revertants in S. typhimurium and E. coli WP2uvrA strains in the presence or absence of S9 mix, compared to the controls. Similarly, low concentrations of syringaresinol (750 and 1,500 microg/plate) did not show any mutagenic properties in all bacterial strains, in the presence or absence of S9 mixture. However, in the high concentration of syringaresinol (3,000 microg/plate), the number of revertants were increased in TA1535 strains, in the absence of S9 metabolic activation. Therefore, in vivo experiments such as comet assay are needed to further determine the genotoxic/carciogenic potential of syringaresinol isolated from A. koreanum.
Eleutherococcus* ; Biotransformation ; Comet Assay ; Escherichia coli ; Immune System Diseases ; Korea ; Lignans* ; Salmonella typhimurium

OBJECTIVESince glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common hereditary hemolytic erythrocyte enzyme deficiency, most cases have single nucleotide mutations in the coding region, and current test methods for gene mutation have some missed detections, this study aimed to investigate the feasibility of RT-PCR sequencing in the detection of gene mutation in G6PD deficiency.

METHODSAccording to the G6PD/6GPD ratio, 195 children with anemia of unknown cause or who underwent physical examination between August 2013 and July 2014 were classified into G6PD-deficiency group with 130 children (G6PD/6GPD ratio <1.00) and control group with 65 children (G6PD/6GPD ratio≥1.00). The primer design and PCR amplification conditions were optimized, and RT-PCR sequencing was used to analyze the complete coding sequence and verify the genomic DNA sequence in the two groups.

RESULTSIn the G6PD-deficiency group, the detection rate of gene mutation was 100% and 13 missense mutations were detected, including one new mutation. In the control group, no missense mutation was detected in 28 boys; 13 heterozygous missense mutations, 1 homozygous same-sense mutation (C1191T) which had not been reported in China and abroad, and 14 single nucleotide polymorphisms of C1311T were detected in 37 girls. The control group showed a high rate of missed detection of G6PD deficiency (carriers) in the specimens from girls (35%, 13/37).

CONCLUSIONSRT-PCR sequencing has a high detection rate of G6PD gene mutation and a certain value in clinical diagnosis of G6PD deficiency.

Adolescent ; Child ; Child, Preschool ; Female ; Glucosephosphate Dehydrogenase ; genetics ; Glucosephosphate Dehydrogenase Deficiency ; diagnosis ; genetics ; Humans ; Infant ; Male ; Mutation ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA

Objective To explore clinical and muscular pathological features of statin-induced myopathy.Methods Nine patients were enrolled in this study,who were diagnosed as statin-induced myopathy by muscle biopsy in Peking University First Hospital from April,2012 to October,2014.The clinical data and pathological findings were analyzed.Results The exposure time to statins varied from 4 days to 4 years in the total of 9 patients,6 males and 3 females,with the average age of 63 ± 6 (55 to 74) years old.Three patients suffered from myalgia and 6 patients complained of weakness mainly at the proximal limbs,while no symptoms occured in 3 patients.Serum creatine kinase (CK) increased in all patients with the maximum value varied from 468 to 8 000 U/L.Serum myositis antibodies were tested in 7 patients and all were negative.Electromyogram was performed in six patients with myogenic damage found in 2 patients.MRI of bilateral thigh muscle was carried out in six patients with muscle edema and mild fatty infiltration found in 2 patients.All patients underwent skeletal muscle biopsy with histochemical and immunohistochemical staining.The main muscular pathological features were muscle fiber atrophy,necrosis,regeneration and increased lipid droplets.Ragged blue fiber,cytochrome C oxidase-negative muscle fibers and decreased NADH activity were observed in some patients.MHC-Ⅰ expressed in the sarcolemma of muscle fibers at various levels.Mild C5b-9 staining was found in the endomysium,capillary and cytoplasm.Symptoms and the level of CK were improved in 7 patients after discontinuing statins or changing to another statin,while the immunosuppressive therapy were used in 2 patients and shown to be effective.Conclusions Statin induced myopathy is self-limiting in most patients,with improvement after discontinuation of statins.Few patients with autoimmune necrotic myopathy need immunosuppressive therapy.

Objective To analyze the clinical,myopathological and genetic features in 5 female manifesting carriers of Duchenne muscular dystrophy (DMD).Methods The age of onset of these 5 patients were from birth to 54 years old,one of which had a family history of DMD.Two patients presented with proximal weakness,one with myalgia and dilated cardiomyopathy,one with limb weakness and ventricular septal defect,and one with exercise intolerance.Serum creatine kinase concentrations were between 1 000-31 815 U/L.Muscle biopsies were performed in 4 patients.Dystrophin gene mutation analyses were carried out in 5 patients by multiplex ligation-dependent probe amplification.Karyotype study was done in one patient who had no dystrophin gene mutation.Results Muscle biopsy revealed markedly decreased dystrophin expression in one patient and a mosaic pattern with some fibers lacking or partially expressing dystrophin in 3 patients.Four patients were identified carrying exonic deletions of dystrophin gene and one had t(x;5) (p21 ;p14).Conclusions The clinical manifestations and myopathological changes are more compatible with Becker muscular dystrophy.Chromosome translocation can be detected in Chinese female manifesting carrier.

Objective To investigate the in vitro effects of quercitrin on the proliferation and the cytotoxicity of human γδT cells.Methods Peripheral blood mononuclear cells (PBMCs) were isolated from healthy subjects and cultured with isopentenyl pyrophosphate and IL -2 to induce human γδT cells.The hu-manγδT cells were cultured with quercitrin at various concentrations for 48 hours.CCK-8 kits were used to analyze the in vitro proliferation and cytotoxic activities of γδT cells.Flow cytometry was performed to meas-ure the expression of granzyme B and perforin in γδT cells.The expression of p-ERK, p-Akt and Bcl-2 at protein level were detected by Western blot .Results The percentage of human γδT cells in PBMCs was in-creased from (2.96±1.83)%to (88.94±2.36)%after 10 days of culture.The quercitrin at concentrations of 10 to 80 μg/ml could promote the growth of γδT cells and up-regulate the expression of granzyme B , per-forin, p-ERK, p-Akt and Bcl-2 in a dose dependent manner .The cytolytic activities of γδT cells against co-lonic carcinoma cells ( HCT116 ) were enhanced by quercitrin .Conclusion Quercitrin could promote the proliferation of γδT cells and enhance the expression of granzyme B and perforin at certain concentrations in vitro.ERK1/2 and Akt signal transduction systems might be involved in the process .

Objective To evaluate the value of urine neutrophil gelatinase-associated lipocalin (uNGAL) to early diagnose acute kidney injury(AKI) of critically ill children in PICU.Methods Eighty critically ill children at PICU of Children's Hospital Affiliated to Shanghai Jiaotong University were enrolled in this study from April to June 2013.They were continuously observed for 72 hours.According to pediatric RIFLE criteria for diagnosis of AKI,patients were divided into AKI group (15 cases) or non-AKI group (65 cases).Additionally,according to sepsis diagnostic criteria,patients were divided into sepsis group (31 cases) or non-sepsis group (49 cases).The levels of serum creatinine and uNGAL were measured within 6th hour,24th hour,48th hour,72th hour after admitted to PICU.The differences of uNGAL levels between AKI and non-AKI groups,sepsis without AKI and non-sepsis non-AKI groups,sepsis merged AKI and sepsis without AKI groups were analysed.The sensitivity and specificity of uNGAL and serum creatinine for diagnosis of AKI at 48th hour were evaluated by ROC curve.Results Thirteen cases of eighty children developed to AKI after admitted to PICU.(1)The uNGAL levels [M(QR),ng/ml] in AKI group within 6th hour,at 24th hour,48th hour,72th hour were 863.00 (696.00),700.50 (580.00),365.50 (285.00),289.50 (319.30),respectively,which were significantly higher than those in non-AKI group [20.00 (106.00),20.00 (85.30),20.00(101.00),20.00(36.00)] (P ＜0.01).(2)The uNGAL levels in new developed group were much higher than those in non-AKI group at each time point.The comparision of serum creatinine at 48th hour was statistic difference.(3)The uNGAL levels rised at early stage in sepsis without AKI group and down to normal gradually after 48th hour.(4)The uNGAL levels continued increasing in sepsis merged AKI group,and had significant differences comparing with sepsis without AKI group(P ＜ 0.01).(5) The areas under ROC curve of uNGAL and serum creatinine at 48th hour were 0.902(95％ CI:0.801 ～ 1.004) and 0.801 (95％ CI:0.768 ～ 0.981),respectively.Conclusion The level of uNGAL has earlier increase for 24 to 48 hours than that of serum creatinine in critically ill children,and it can also reflect the severity of AKI.Therefore it can be used as an early diagnostic biomarker for AKI in PICU.