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.

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

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

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

Objective To explore the relationship of expression and the effect of preoperative radiotherapy of endothelin-1 (ET-1) and pyruvate kinase M-2 (PKM2) in rectal carcinoma.Methods Immunohistochemical method was used to detect the expression of ET-1 and PKM2 proteins of rectal cancer tissues in 96 cases.The expressions of ET-1 and PKM2 were analyzed with the effect of preoperative radiotherapy in rectal cancer tissue.Results The high expression of ET-1 protein was 59 cases (61.46％).The high expression of PKM2 proteins was 54 cases (56.25％).The high expressions of ET-1 and PKM2 protein were worsen the effect of tumor regressive grade (TRG) than lower expressions of those after preoperative radiotherapy of rectal cancer tissue (P ＜ 0.05).The protein expression of ET-1 and PKM2 were positively correlated (P =0.006).Conclusions The high expressed ET-1 and PKM2 proteins in rectal cancer are closely related to preoperative radiotherapy resistance.ET-1 and PKM2 proteins are expected to become new targets of radiotherapy sensitivity and radiotherapy sensitization of rectal cancer.

Objective To explore the protective effect of rnicroRNA (miRNA)-155 inhibitor on interleukin-1 receptor-associated kinase (IRAK)-1 mRNA and IRAK-4 mRNA in endotoximia induced liver injury in mice.Methods One hundred and twenty male BALB/c mice were randomly divided into healthy control group(n =40),endotoximia group (n =40) and miRNA-155 inhibitor group (n =40).Each group were divided into 6 h,12 h,24 h,48 h subgroups,each of which consisted of 10 mice.The mice in miRNA-155 inhibitor group were administered with miRNA-155 inhibitor[80 mg(kg ·d)] via tail vein injection before lipopolysaccharide (LPS) administration while the other two groups treated with normal saline,following 24 hours,model of endotoximia mice was produced by injection of LPS intraperitoneally.At 6 h,12 h,24 h,48 h after LPS exposure,the experimental mice were sacrificed and the liver tissue samples were collected.Histopathological changes,the expression of miRNA-155,IRAK-1 mRNA,IRAK-4 mRNA,tumor necrosis factor (TNF)-α,IL-1,IL-10 were detected.Results LPS exposure resulted in increase of miRNA-155,IRAK-1 mRNA,IRAK-4 mRNA,TNF-α,IL-1 and IL-10 in both endotoximia group and miRNA-155 inhibitor group compared to the control group,miRNA-155 inhibitor resulted in decrease of miRNA-155,IRAK-1 mRNA,IRAK-4 mRNA,TNF-α,IL-1 and IL-10 in miRNA-155 inhibitor group compared to the endotoximia group.There were significant differences of miRNA-155 expression at 12 h,24 h,48 h after LPS exposure among 3 groups (P ＜ 0.05).Both IRAK-1 mRNA and IRAK-4 mRNA showed significant differences at 12 h,24 h,48 h.Turning to inflammation factors,differences were found among 3 groups at all time points (P ＜ 0.05).At light-scope,there was improvement in sepsis associated liver injury in miRNA-155 inhibitor group compared to endotoximia group.Conclusion miRNA-155 inhibitor administration appears to down regulate IRAK-1 mRNA and IRAK-4 mRNA expression and further deduce the excessive inflammatory and anti-inflammatory reaction,which may alleviate liver injury in endotoximia mice.

Objective To report the clinical and peripheral neuropathological findings in two patients with autosomal recessive Charcot-Marie-Tooth disease 2K(AR-CMT2K).Methods Case one was a nine year-old girl.She had distal weakness of lower limbs for six years, with calf atrophy and contracture of Achilles tendon for three years.Case two was an eight year-old boy.He had distal weakness of lower limbs with contracture of Achilles tendon and calf muscle atrophy for three years, and proximal weakness of low limbs for two years.The motor nerve conduction velocities in median nerves were 48.1 m/s in case one and 47.6 m/s in case two.The compound motor action potential amplitude of median nerves decreased by 46% in case one and 69% in case two.Sural nerve biopsies and gene targeted next-generation sequencing were performed in both patients.Results Density of myelinated fibers was 8 407/mm2 in case one and 7 714/mm2 in case two.The ratio of myelinated fibers with diameter over 8 μm was 2.6% in case one and 0 in case two.Both patients had small regenerating cluster of myelinated fibers.Thin myelinated fibers appeared in case one.In case two, atypical onion bulb formations with focal folded myelin appeared, and electromicroscopy revealed mitochondrial aggregate in axons.Compound heterozygous mutations of ganglioside-induced differentiation associated protein 1 gene were detected in both patients, including c.767A>G(p.H256R) and c.466G>A (p.A156T) in case one and c.767A>G and 845G>A(p.R282H) in case two.Conclusions Contracture of Achilles tendon may appear in early childhood of AR-CMT2K patients.The main pathological changes in sural nerve are loss of large myelinated fibers, mitochondrial aggregate in axons and myelin abnormalities.

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 .