Objective To investigate the association between polymorphism of HLA-DRB1 allele and systemic sclerosis (SSc) and scleroderma-associated renal damage in Han Chinese of Henan Province.Methods Eighty-one SSc patients and 90 normal controls were recruited in this study,among them 59 patients had renal damage (SRD).The genotyping was carried out by nest PCR-SBT and gene clone.Comparisons between groups were performed with x2 test or exact probabilities.Multivariable Logistic regression was used to evaluate the association between prevalence of SSc or SRD and the possible relevant alleles.Results Thirty-three HLA-DRB1 alleles were discovered from the specimens,including 27 in SSc specimens,and 22 in SRD.Among them,the allele frequencies of HLA-DRB1 * 040501 (8.64％), * 110101 (11.11％), * 150201(8.02％) were higher in SSc patients than those of the controls (1.67％,4.44％,2.22％ respectively).After adjusted for other factors,HLA-DRBl * 040501 (P=0.010,OR =5.839,95％CI:1.518-22.460)、* 110101(P=0.019,OR=3.060,95％CI:1.204-7.772)、* 150201(P=0.010,OR=4.780,95％CI:1.444-15.821 )were identified as independent risk factors for SSc.And the allele frequencies of HLA-DRB1*040501 (9.32％),* 150201 (7.63％) were higher in SRD patients than those of the controls (1.67％,2.22％ respectively).After adjusted for other factors,HLA-DRB1 * 040501 (P=0.008,OR=6.363,95％CI:1.614-25.087) and * 150201 (P=0.030,OR =4.043,95 ％CI:1.147-14.243 ) were identified as independent risk factors for SRD.Conclusion Our data suggest that HLA-DRB1 * 040501,* 110101,* 150201 may be susceptible genes for SSc and the HLA-DRB1 * 040501,* 150201 may be susceptible genes for SRD in Han Chinese of Henan Province.

Objective To detect the serum level of mannose binding lectin (MBL) in patients with renal injury induced by rheumatoid arthritis (RA), and to investigate the role of MBL in the pathogenesis of renal injury in RA. Methods ELISA was used to measure the serum MBL level of 19 RA patients with renal injury, 49 RA patients without renal injury and 40 healthy individuals. The clinical features and laboratory markers were compared and analyzed by chi-square test, two independent samples t-test and Spearman's correlation analysis. Results Compared with RA patients without renal injury, the number of tender and swollen joints [(15±9) vs (9±11)], duration of morning stiffness [(2.9±1.3) vs (2.3±1.6) h] and extraarticular manifestations (42.1％ vs 16.3％) in RA patients with renal injury were significantly higher (P＜0.05or P＜0.01). There was no significant difference between the two groups in RA disease duration and jointdeformity(P＞0.05). In patients with renal injury, the level of platelet count [(376±155)×109/L vs (304±121)×109/L], CIC[(4.3±3.0) vs (2.9±3.3) g/L], ESR[(79±46) vs (53±31) mm/1 h], RF[(77±42) vs (52±49)U/ml], CRP[(32±28)vs (23±18)mg/L], IgG[(11.7±2.6)vs (8.4±2.4)g/L], C3[(1.18±0.53)vs (0.94±0.21) g/L] were higher than those in RA patients without renal injury (P＜0.01 or P＜0.05); the level of Alb [(26±13) vs (30±9) g/L] was lower than that in RA patients without renal injury (P＜0.05). The level of serum MBL in the two groups of RA patients was significantly lower than that in the healthy group [(3.1±0.5)mg/L](P＜0.01), and the level of serum MBL in RA patients with renal injury [(1.7±1.2) mg/L] was higher than that in RA patients without renal injury [(1.4±1.3) mg/L](P＜0.05). The level of serum MBL in RA patients with renal injury showed a positive correlation with IgG, C3, CRP and 24 h urine protein level (r=0.6, 0.6, 0.47, 0.57; P＜0.05). Conclusion Renal injury in RA patients is immune complex dependent. The serum level of MBL is higher in patients with renal injury, therefore, high-concentration MBL may be one of a potential causes of renal injury in RA patients.

The genome of CK/CH/SD09/005, an isolate of infectious bronchitis virus (IBV), was characterized to enable the further understanding of the epidemiology and evolution of IBV in China. Twenty-five pairs of primers were designed to amplify the full-length genome of CK/CH/SD09/005. The nucleotide sequence of CK/CH/SD09/005 was compared with reference IBV strains retrieved from GenBank. The phylogenic relationship between CK/CH/SD09/005 and the reference strains was analyzed based on S1 gene sequences. The complete genome of CK/CH/SD09/005 consisted of 27691 nucleotides (nt), excluding the 5' cap and 3' poly A tail. The whole-genome of CK/CH/SD09/005 shared 97 - 99% nucleotide sequence homology with the GX-NN09032 strain, which was the only complete genome that was closely related to CK/CH/SD09/005. When compared with all reference strains except GX-NN09032, CK/CH/SD09/005 showed the highest similarity to ck/CH/LDL/091022 and SDIB821/2012 (QX-like) in the replicase gene (Gene 1) and 3'UTR, with a sequence identity rate of 97% and 98%, respectively. However, CK/CH/SD09/005 exhibited lower levels of similarity with ck/CH/LDL/091022 and SDIB821/2012 in S-3a-3b-3c/ E-M-5a-5b-N with a sequence identity of 72% - 90%. CK/CH/SD09/005 showed the highest level of nucleotide identity with Korean strain 1011, and Chinese strains CK/CH/LXJ/02I, DK/CH/HN/ZZ2004 and YX10, in ORF 3c/E (97%), 5a (96%), 5b (99%) and N (96%), respectively. ORFs 3a, 3b and M of CK/CH/SD09/005 exhibited no more than 90% homology with the reference strains, excluding GX-NN09032. The phylogenic analysis based on the S1 gene revealed that CK/CH/SD09/005 and 39 published strains were classified into seven clades (genotypes). CK/CH/SD09/005 was distributed in clade IV with several isolates collected between 2007 and 2012. CK/CH/SD09/005 showed 66% - 69% and 72% - 81% nucleotide identities with the IBV strains of other six clades in the S1 and S2 subunits, respectively. More over, multiple substitutions were found throughout the entire S gene of CK/CH/SD09/005, while insertions and deletions were located within the S1 gene. These results indicated that CK/CH/SD09/005 is a novel variant that may be derived from the QX-like strains that are prevalent in China. Multiple genetic mechanisms, including recombinations, mutations, insertions and deletions, are likely to have contributed to the emergence of this IBV strain.
Animals ; Chickens ; China ; Coronavirus Infections ; veterinary ; virology ; Genome, Viral ; Genomics ; Infectious bronchitis virus ; classification ; genetics ; isolation & purification ; Molecular Sequence Data ; Phylogeny ; Poultry Diseases ; virology ; Sequence Homology, Amino Acid ; Viral Proteins ; chemistry ; genetics

This study was purposed to develop a real-time PCR assay for sensitive quantification of JAK2V617F allele burden in peripheral blood and to evaluate the clinical value of this method. Both allele-specific mutant reverse primer and wild-type TaqMan-MGB probe were used for dual-inhibiting amplification of wild-type alleles in a real-time PCR, and then the JAK2V617F mutant alleles were amplified specially. The standard curve for quantification of JAK2V617F was established by percentages of JAK2V617F alleles with threshold cycle (Ct) values in a real-time PCR. Furthermore, 89 apparent healthy donors were tested by this method. The results showed that the quantitative lower limit of this method for JAK2V617F was 0.1%, and the intra- and inter-assay average variability for quantifying percentage of JAK2V617F in total DNA was 4.1% and 6.1%, respectively. Two JAK2V617F-positive individuals were identified (the percentage of JAK2V617F alleles were 0.64% and 0.98%, respectively) using this method in blood from 89 apparently healthy donors. It is concluded that the developed method with highly sensitive and reproducible quantification of JAK2V617F mutant burden can be used clinically for diagnosis and evaluation of disease prognosis and efficacy of therapy in patients with myeloproliferative neoplasms. Moreover, this technique can be also used for quantitative detection of variety of single nucleotide mutation.
Adult ; Aged ; Alleles ; Case-Control Studies ; DNA Mutational Analysis ; DNA Primers ; genetics ; Genotype ; Humans ; Janus Kinase 2 ; genetics ; Middle Aged ; Mutation ; Myeloproliferative Disorders ; genetics ; Real-Time Polymerase Chain Reaction

OBJECTIVETo investigate the neurodevelopmental outcome of preterm infants who were discharged from neonatal intensive care unit (NICU) at 1 year of age and the impact factors contributing to the neurodevelopmental outcome and to study whether early intervention can improve the neurodevelopmental outcome in preterm infants.

METHODSEarly intervention guidance and follow-up visits were performed after the preterm infants discharged from NICU. The infants received the infant development test of Child Development Centre of China (CDCC) for neurological development at 1 year of age. The neurodevelopmental outcome was evaluated based on mental development index (MDI) and physical development index (PDI). MDI and PDI > 85 was defined as normal, MDI or PDI between 70 and 85 as critical and MDI or PDI < 70 as abnormal. Social-domestic and clinical factors related to neurological development were analyzed by ANOVA or chi-square test. Potential high risk factors were analyzed with logistic regression. To investigate the effects of intervention compliance on neurological development, the infants were classified into two groups according to different intervention compliances. The good compliance group included infants who received a through familial intervention for more than 4 days weekly (30 minutes daily) and consulted with physicians more than 5 times in 1 year. The infants who did not receive the interventions as the good compliance group served as the bad compliance group.

RESULTSThis study consisted of 210 infants, with a mean gestational age of 33.2 +/- 2.6 weeks and a mean birth weight of 1923.3 +/- 558.8 g. Normal, critical and abnormal neurological development occurred in 123 cases (58.6%), 61 cases (29.0%) and 26 cases (12.4%) respectively. Eighteen infants (8.6%) had mental lag and 9 (4.3%) had cerebral palsy (CP). The MDI and PDI scores of the good compliance group (111 cases) were 97.15 +/- 17.38 and 94.23 +/- 18.55 respectively, which were markedly higher than those of the bad compliance group (89.87 +/-18.92 and 87.20 +/-19.12; P < 0.05). The incidence of CP (3/111, 2.7%) in the good compliance group was lower than that of the bad compliance group (5/99, 6.1%) although there were no statistical differences. Parents' education level, multiple birth, serious intracranial hemorrhage and apnea were risk factors for adverse neurodevelopmental outcome.

CONCLUSIONSPreterm infants discharged from NICU are a high risk group of neurodevelopmental disablement. Early intervention can improve the neurodevelopmental outcome of perterm infants at high risk.

Child Development ; Humans ; Infant ; Infant, Newborn ; Infant, Premature ; Intensive Care Units, Neonatal ; Logistic Models ; Nervous System ; growth & development ; Patient Compliance ; Prognosis ; Risk Factors

OBJECTIVETo explore the clinical and physical factors that might give rise to radiation-induced esophagitis in three-dimensional conformal radiotherapy for non-small cell lung cancer.

METHODSTo collect the clinical and physical records and follow-up information of 106 NSCLC patients without undergoing surgery in our hospital. χ(2) test, linear tendency test and analysis of variance were employed to analyze the relationship between occurrence of radiation-induced esophagitis and clinical and physical treatment. Logistic analysis was also used for multivariate analysis.

RESULTSAmong the 47 cases of radiation-induced esophagitis, 31 cases were of grade I, 11 of grade II, 5 of grade III, and with a total occurrence rate of 44.3% (47/106). Radiation-induced esophagitis was correlated with Karnofsky scores, radiation sensitization and tumor location (χ(2) = 11.30, 8.45, 7.67, P < 0.05). Radiation-induced esophagitis was correlated with the length of irradiated esophagus and average dose of irradiated esophagus (F = 20.82, 83.08, P < 0.001). With the increase of the irradiated volume percentage from V20, V30, V40 up to V50, the occurrence rate of radiation-induced esophagitis was also increased, almost with a linear trend (P < 0.05). Application of all the above factors to logistic model indicated that radiation sensitization,length of irradiated esophagus, average dose and V50 were all statistically significant foactors in the occurrence of radiation-induced esophagitis (OR = 0.321, 2.850, 7.307 and 8.558, P < 0.05).

CONCLUSIONSRadiation sensitization,length of irradiated esophagus, average dose of irradiated esophagus and V50 are independent factors in the occurrence of radiation-induced esophagitis. V50 is of greater importance in the judgement of occurrence of radiation-induced esophagitis.

Aged ; Carcinoma, Non-Small-Cell Lung ; radiotherapy ; Esophagitis ; etiology ; Esophagus ; pathology ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; radiotherapy ; Male ; Middle Aged ; Multivariate Analysis ; Radiation Injuries ; etiology ; Radiation Tolerance ; Radiotherapy Dosage ; Radiotherapy, Conformal ; adverse effects ; methods ; Retrospective Studies

OBJECTIVETo develop a novel real-time PCR for sensitively quantitative detection of JAK2 V617F allele burden in peripheral blood.

METHODSBased on the real-time allele-specific PCR (AS-qPCR), the locked nucleic acid (LNA)-modified oligonucleotide probe was used for selectively blocking amplification of wild-type alleles in AS-qPCR, and then a novel AS-LNA-qPCR method was established. The percentages of sample JAK2 V617F alleles were directly calculated by its threshold cycle (Ct) values according to the standard curve which generated by JAK2 V617F alleles with its Ct values. We validated intra- and inter-assay variability for quantifying JAK2 V617F. We also assayed 623 apparent healthy donors by our method to validate its clinical application value.

RESULTSThe quantitative lower limit of this method for JAK2 V617F was 0.01%, and the intra- and inter-assay average variability for quantifying percentage of JAK2 V617F in total DNA was 6.3% and 8.6%, respectively. Nineteen JAK2 V617F-positive individuals were identified using AS-LNA-qPCR in blood of 623 apparently healthy donors, and the range of percentages of JAK2 V617F alleles were 0.01%-5.49%.

CONCLUSIONThe AS-LNA-qPCR with highly sensitive and reproducible quantification of JAK2 V617F mutant burden can be used clinically for diagnosis as well as evaluation of disease prognosis and efficacy of therapy in patients with myeloproliferative neoplasms.

Alleles ; Humans ; Janus Kinase 2 ; genetics ; Mutation ; Oligonucleotide Probes ; genetics ; Oligonucleotides ; genetics ; Real-Time Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

OBJECTIVETo investigate the expression of epidermal growth factor receptor (EGFR) during the mineralization of human periodontal ligament cells (hPDLC) in vitro.

METHODSStudies using specific antibodies to immunolocalize EGFR in the mineral differentiating hPDLC were undertaken to investigate the different expression during the inducing process. In situ hybridization and RT-PCR technique were used to investigate the transcripts encoding the protein of EGFR.

RESULTSThe results showed that immunocytochemical labeling gradually decreased following the elong of the induce time, downing to nearly negative at the 4th week and the signal of EGFR transcripts was weaker in the induced hPDLC than that in uninduced.

CONCLUSIONEGFR has a negative regulation function during the mineralization of hPDLC.

Cells, Cultured ; Humans ; In Situ Hybridization ; In Vitro Techniques ; Periodontal Ligament ; Receptor, Epidermal Growth Factor

The study was purposed to investigate whether the cyclooxygenase inhibitors from some dietary vegetables can inhibit platelet aggregation function by the arachidonic acid (AA). The vegetable juice was mixed with platelet rich plasma (PRP), and asprin was used as positive control. The maximum ratio of platelet aggregation induced by AA was measured on the aggregometer; heme and cyclooxygenase-1 (COX(1)) or cyclooxygenase-2 (COX(2)) were added to test tubes containing COX reaction buffer, the mixture was vortex-mixed and exposed to aspirin or vegetable juice, followed by addition of AA and then hydrochloric acid (1 mol/L) was added to stop the COX reaction, followed by chemical reduction with stannous chloride solution. The concentration of COX inhibitors was detected by the enzyme immunoassay kit; vegetable juice (aspirin as positive control) was mixed with whole blood, which was followed by the addition of AA, and then the reaction was stopped by adding indomethacin, centrifuged, then the supernatant was collected, and the plasma thromboxane B(2) (TXB(2)) was measured by radioimmunoassay. The results showed that spinach juice, garlic bolt juice, blanched garlic leave juice and Chinese leek juice could inhibit by 80% human platelet aggregation induced by AA. 4 kinds of vegetables were all found a certain amount of cyclooxygenase inhibitors, which COX(1) and COX(2) inhibitor concentrations of spinach were higher than that of aspirin; 4 vegetable juice could significantly reduce the human plasma concentrations of TXB(2) induced by AA (p < 0.05). It is concluded that 4 kinds of raw vegetables containing cyclooxygenase inhibitors inhibit the production of TXA(2) and thus hinder platelet aggregation. Raw spinach, garlic bolt, blanched garlic and chinese leek inhibit significantly AA-induced human platelet aggregation in vitro. 4 kinds of vegetables may have a good potential perspective of anti-platelet aggregation therapy or prevention of thrombosis.
Adult ; Arachidonic Acid ; metabolism ; Blood Platelets ; drug effects ; Cyclooxygenase Inhibitors ; pharmacology ; Female ; Humans ; Male ; Platelet Aggregation ; drug effects ; Vegetables ; chemistry

OBJECTIVETo study the effects of VAC on starting the process of wound healing and decreasing apoptosis.

METHODSTo examine the variations in expression of proto-oncogenes c-myc, c-jun and Bcl-2 in pig wound model with acute full-thickness skin defect and human chronic wounds by immunohistochemistry, calculate the numbers of expressive positive cells and the labelling index (LI), and observe the process of wound healing.

RESULTS(1) In pig experiment, the wound in experimental group was very clean and without obvious exudates, many neoepiderm and granulation tissue rapidly appeared or formed after 6 days, and healed completely by the 25th day. On the contrary, in the wound of control group, more exudates and blood crust could be seen and fewer neoepiderm and granulation tissue appeared after 6 days and was healed by 30th day. Immediately after the wound was created, the expression of c-myc, c-jun and Bcl-2 was lower and mainly situated in nucleus or cytoplasma of the basilar cells. After the wound was created in control group, or after starting the VAC treatment in experimental group, their expression rapidly and obviously increased, the distribution of the positive cells also became enlarged, but the amount of expression decreased rapidly after the expressive peak have reached. In the successive 12 days following the wound was created, the expression of c-myc, c-jun and Bcl-2 in the experimental group was constantly higher than that of the control group. (2) In human chronic wounds, there wasn't obvious secretions and more healthy granulation tissue was rapidly formed after VAC treatment. The expression of c-jun was mainly located in cytoplasma of basilar cells of epithelium, dermal fibroblasts and inflammatory cells, and the positive cell and labelling index obviously decreased. The expression of c-myc and Bcl-2 was mainly in cytoplasma of basilar cells, but the amount of expression and the labelling index became obviously increased after VAC treatment.

CONCLUSIONSVAC could rapidly start the healing course of the pig' s acute skin wound and human chronic wound, decrease apoptosis of the reparative cells, so as to accelerate wound healing.

Adult ; Animals ; Apoptosis ; Female ; Humans ; Male ; Middle Aged ; Negative-Pressure Wound Therapy ; Proto-Oncogene Proteins c-jun ; metabolism ; Proto-Oncogenes ; Swine ; Wound Healing