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.

OBJECTIVETo explore the character of inorganic elements in Asari Radix et Rhizoma (Xixin).

METHODThe contents of 53 inorganic elements in Xixin samples from different localities and species were determined by ICP-AES and ICP-MS. The statistical data were made using SAS.

RESULTThe result demonstrated that Xixin has the high contents of Fe, Cr, Li. It has been observed that the content of Cu and Pb of the samples are much higher than the standard level. The results of hierarchical cluster analysis revealed two groups which correspond with the species of the samples. No correlations between the contents of the inorganic elements and the localities of the samples were found. Some characteristic elements were displayed in some specific areas. The difference of the contents of the 53 inorganic elements between root and rhizome of Xixin was reported for the first time. The primary form of inorganic elements in Xixin has been studied for the first time. The result demonstrated that the extraction rate between different elements varied, with the average extraction rate of (22.25 +/- 24.96)%.

CONCLUSIONThe inorganic elements analysis of Xixin can provide evidence of its identification, cultivation and application.

Asarum ; chemistry ; classification ; China ; Drugs, Chinese Herbal ; analysis ; Rhizome ; chemistry ; Trace Elements ; analysis

AIMTo immunolocalize the c-mos gene product and to investigate its spatial and temporal expression in mouse testis during postnatal development.

METHODSSemi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization techniques were used to examine c-mos mRNA and indirect immunofluorescence was used to localize c-Mos protein in mouse testis on postnatal days 14, 21, 25, 28, 30, 35, 49 and 70.

RESULTSc-mos mRNA remained low on postnatal days 14-21, increased abruptly from day 25 and peaked on day 30. Its levels decreased a little on day 35 and became almost stable thereafter until day 70. c-mos mRNA was localized in the nucleus and cytoplasm of the spermatocytes and round spermatids. The nuclear staining was much stronger than the cytoplasmic staining. Using a polyclonal anti-c-Mos antibody, Western blotting detected a single band at 43 kDa in testis lysate. c-Mos protein was exclusively localized to the elongating spermatids and was first detected on postnatal day 30. The number of c-Mos-positive spermatids increased progressively till day 49 and stabilized thereafter.

CONCLUSIONThe c-mos gene displays a spatial and temporal expression pattern in the mouse testis during postnatal development at both the mRNA and protein level. This suggests that c-mos might play important roles in spermatogenesis.

Animals ; Fluorescent Antibody Technique, Indirect ; Gene Expression ; Genes, mos ; genetics ; Male ; Mice ; Reverse Transcriptase Polymerase Chain Reaction ; Spermatocytes ; growth & development ; metabolism ; Spermatogenesis ; genetics ; Testis ; metabolism

OBJECTIVETo investigate the clinical effects and prevent the complications of posterior and anterior decompression and internal fixation in the revision of cervical anterior internal fixation failure.

METHODSFrom 2008 January to 2011 December, 17 patients with cervical anterior internal fixation failure were treated with posterior and anterior decompression and internal fixation. There were 12 males and 5 females, aged from 26 to 68 years old with an average of 44.1 years. The lower screw loosening was found in 6 cases, the upper screw loosening in 5 cases, titanium mesh caving in 3 cases, the upper screw breakage in 2 cases, the lower screw breakage in 1 case. Informations of bone fusion were observed by X-ray, CT, MRI. Clinical effects were evaluated by modified JOA score.

RESULTSAll the revision operations were successfully completed. One case with poor blood coagulation function before operation resulted in postoperative hematoma and occurred neurological symptoms; after hematoma removal and fresh frozen plasma infusion later, neurological symptoms of the patient disappeared. All patients were followed up from 6 to 38 months with an average of (22.4±10.0) months. Postoperative at 2 weeks, 3 months, and final follow-up, JOA score had obviously improved and respectively was 13.1±1.6, 13.4±1.6, 14.2±1.5. All internal fixation locations were good after revision,and obtained bone fusion at 10 months after operation, with an average fusion time of 6 months.

CONCLUSIONThe combined posterior and anterior decompression and internal fixation in the revision of cervical anterior internal fixation failure is safe, can achieve thoroughly decompression, maintain the cervical curvature, reconstruct the three column stability, and it may be used for the patients of cervical anterior fixation failure.

Adult ; Aged ; Cervical Vertebrae ; surgery ; Decompression, Surgical ; methods ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged

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 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

BACKGROUNDMycoplasma pneumoniae (M. pneumoniae) is one of the common pathogens causing atypical pneumonia. In recent years, resistance to macrolides has become more common, especially in China. Previous studies have confirmed that the mutation at position 2063 in domain V of the 23S rRNA is the most prevalent, followed by the mutation at position 2064. Reported molecular detection methods for the identification of these mutations include direct sequencing, restriction fragment length polymorphism analysis, real-time polymerase chain reaction (PCR) with high-resolution melt analysis, and nested PCR-linked with capillary electrophoresis, etc. The most commonly used method for monitoring resistance-conferring mutations in M. pneumoniae is direct DNA sequencing of PCR or nested PCR products. However, these methods are time-consuming, labor-intensive or need expensive equipments. Therefore the development of rapid and sensitive methods is very important for monitoring the resistance globally.

METHODSIn this study, we reported a fast and cost-effective method for detecting 2063 and/or 2064 macrolide resistant mutations from specimens using a modified allele-specific PCR analysis, and all results were compared with the sequencing data. We also analyzed the clinical courses of these samples to confirm the modified allele-specific PCR results.

RESULTSAmong 97 M. pneumoniae specimens, 88 were found to possess mutations by this method, and all modified allele-specific PCR analysis results were consistent with the sequencing data. The data of the clinical courses of these 97 cases showed that they suffered from severe pneumonia. Erythromycin showed better efficacy on cases from which no macrolide resistance mutation was found on their specimens. However, in some cases from which mutations were detected, erythromycin monotherapy had poor efficacy, and on these patients severe symptoms improved only when azithromycin was added to the treatment.

CONCLUSIONSThe drug-resistant M. pneumoniae is very common in Beijing, China. Our modified allele-specific PCR analysis can identify erythromycin resistant mutations more rapidly from specimens than any other method currently available. Erythromycin is still effective for treating patients infected with the mutation negative M. pneumoniae, but this treatment fails to work on mutant organisms. This method can facilitate clinicians in selecting appropriate therapy within short timescales.

Alleles ; Anti-Bacterial Agents ; pharmacology ; China ; Drug Resistance, Bacterial ; genetics ; Erythromycin ; pharmacology ; Mycoplasma pneumoniae ; drug effects ; genetics ; Polymerase Chain Reaction ; methods

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

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