Objective To establish matched-pairs of DNA samples with copy-number controlled differential target genes, and to compare the detection sensitivity of typical Representational Difference Analysis (RDA) method and Subtractive Hybridization Difference Display (SHDD) method in isolating differential genes.Methods Two gene fragments (376 bp and 869 bp in length respectively) cloned by PCR using Human Papilloma Virus (HPV) DNA as template were used as differential target genes, and mixed with human genome DNA. Five matched, pairs of human genome DNA samples with gradually increased difference in copy numbers of target genes were established and RDA and SHDD methods were performed to clone differential target genes and compared their detection sensitivity. Results By using RDA method, 376 bp fragment with 6-fold difference and 869 bp fragment with 8-fold difference were cloned.However, both of these two target fragments with 4-fold difference were isolated using SHDD method.Conclusion The SHDD method adopts balanced bi-directional subtractive hybridization between two sample difference products and avoids loss of differential target genes caused by unbalanced subtractive hybridization of RDA method, and thus outweighs RDA method in isolating target genes, especially long-length target fragments, with small difference.

Heterogeneous nuclear ribonucleoprotein (hnRNP) is a family of multifunctional nuclear RNA-binding proteins that regulate the alternative splicing of pre-mRNA and the transport, translation, and stability of mRNA. The most abundant and best charac-terized proteins of this group are hnRNP A1 and hnRNP A2, which share a high degree of sequence homology and functional similarity. HnRNP A1 and hnRNP A2 are upregulated in multiple human tumors and modulate the alternative splicing and mRNA stability of vari-ous tumor-related genes critical to tumor cell growth, apoptosis, inflammatory and immune reactions, and epithelial-to-mesenchymal transition. Therefore, hnRNP A1 and hnRNP A2 have potential diagnostic, prognostic, and therapeutic values.

Objective: To explore the efficacy of mild moxibustion plus loratadine tablets for children with allergic rhinitis (AR).Methods: A total of 80 children were randomized into a control group and an observation group, with 40 cases in each group. The control group was treated with loratadine tablets, and the observation group was treated with mild moxibustion plus loratadine tablets. Before and after treatment, the total nasal symptom score (TNSS) was evaluated, and the serum eosinophils (EOS) count, and the interleukin (IL)-27 and macrophage migration inhibitory factor (MIF) levels were measured. Clinical efficacy was evaluated after treatment. Results: The total effective rate of the observation group was higher than that of the control group (P<0.05). After treatment, the TNSS in both groups decreased (P<0.05), and the TNSS in the observation group was lower than that in the control group (P<0.05); the serum EOS count in both groups decreased (P<0.05), and the serum EOS count in the observation group was lower than that in the control group (P<0.05). The serum IL-27 level in the control group had no statistical difference compared with the same group before treatment (P>0.05), and the serum MIF level decreased after treatment (P<0.05). The serum IL-27 level in the observation group increased after treatment (P<0.05), and the serum MIF level decreased after treatment (P<0.05), and were both statistically different from those in the control group (P<0.05). Conclusion: Mild moxibustion plus loratadine tablets is effective in treating children with AR. It can significantly improve the nasal symptoms and reduce the serum EOS count, which may be related to the regulation of the serum IL-27 and MIF levels.

Objective To investigate tumor lymphatic and mircovascular densities as prognostic markers in 69 cases of invasive breast cancer treated with partial or total mastectomy and lymph node dissection.Methods 69 cases of untreated primary unilateral invasive ductal breast carcinomas were selected.All cases were immunostained with D2-40 and CD31.Positively stained microvessels were counted in densely vascular/lymphatic foci (hot spots).The relationship between lymphatic vessel density (LVD),microvessel density(MVD) and prognosis was analyzed.Results The mean ± SD peritumoral lymphatic vessel density (P-LVD) was significantly higher than intratumoral LVD(I-LVD) (P ＜ 0.01).There was a positive correlation of D2-40 LVD(peritumoral) counts with lymph node metastasis (P =0.003) and clinical stage (P =0.026),and CD31 microvessel density was found significantly associated with clinical stage(P =0.038).No significant association was found between above variants with I-LVD (P ＞ 0.05).Univariate analysis showed that survival time was impaired by higher MVD and higher peritumoral LVD(P =0.007,P =0.008,P =0.014,P =0.024,log-rank test),but not I-LVD.Multivariate survival analysis showed that MVD,peritumoral LVD,TNM stage and lymph node metastasis were independent prognostic factors for overall survival.Conclusions Peritumoral LVD and MVD were significantly correlated with survival status of patients with breast cancer.This is the first attempt to predict prognosis of breast cancer patients by quantifying the peritumoral LVD and MVD.

Objective To prepare the small intestinal submucosa (SIS) to be used as a scaffold in constructing engineered cardiac tissue. Methods Small intestinal submucosa of pig was treated with 0.25% trypsin and then 0.5% SDS for 24 hours each to obtain decellularized SIS. The obtained SIS was tested for its mechanical strength by stretching both horizontally and longitudinally with a material test machine. The cytotoxicity and histocompatibility of the material was also examined. Then the cardiomyocytes harvested from 3-day old neonatal rats were seeded on the SIS to construct engineered cardiac tissue sheets. These acquired engineered cardiac tissue sheets were immunohistochemically stained and observed with inversion microscope and transmission electromicroscope to evaluate their characteristics. Results SIS was decellularized completely. The mechanical capability of the decellularized SIS was satisfactory. Under 15% stretching, its strain and stress was nearly linear. SIS showed no cytotoxicity and inflammatory response. After 12 hours, the cardiomyocytes seeded in the SIS scaffold began to beat spontaneously. Two days later, the cardiomyocytes-SIS scaffold composite (engineered cardiac tissue sheet) began to beat spontaneously, and could beat spontaneously for 14 days. The constructed engineered cardiac tissue sheets consisted of layers of cardiomyocytes and with abundant extracellular collagen in the sheet. Conclusions SIS with good mechanical capability and biocompatibility has been prepared successfully, and then the engineered cardiac tissue has been constructed successfully based on the SIS scaffold.

Objective To observe the long term results of intertrochanteric varus medial displacement osteotomy(IVMDO) for Perthes disease. Methods Thirty eight patients with Perthes disease treated with IVMDO were reviewed. The results were evaluated based on a criteria made by the authors including clinical and radiographic parameters. The duration of follow up ranged from 3 to 15 years, with an average of 7 years. Results Fifteen patients were evaluated as having excellent result, 17 good, 3 fair and 3 poor respectively. The overall excellent or good rate was 84.2%. Considering the relationship between the outcome and staging of the disease, the overall excellent or good rate was 94.7% in stage Ⅱ lesion, 85.7%in stage Ⅲ lesion, and 40.0% in stage Ⅳ lesion. Conclusion The treatment of Perthes disease with IVMDO has the advantages of simple manipulation, less trauma and good results, and is worthy of populariztion.

RAPD analysis of eight soy sauce strains isolated from commercial soy sauce koji was done with random primers,using Aspergillus oryzae AS3.951 and Aspergillus sojae AS3.495 as controls.Six appropriate primers(Primer1,Primer5,Primer6,Primer7,Primer8,Primer9)for RAPD-PCR were screened from nine random primers,and repetitive experiments demonstrated that their RAPD-PCR polymorphic patterns were stable.There were usually 4～8 bands in their RADP-PCR patterns,where the number of the main bands was 1～4 and the secondary bands were abundant.The phylogenetic tree of these strains was reconstructed according to their RAPD-PCR patterns,and it basically corresponded to traditional morphological taxology,demonstrating that the application of RAPD molecular marker in the phylogenetic analysis of soy sauce strains was feasible.

OBJECTIVETo explore the effect of Danlou Tablet (DT) on arrhythmia model rats induced by transient myocardial ischemia/reperfusion (I/R).

METHODSTotally 45 healthy Wistar rats were randomly divided into 3 groups, the sham-operation group, the model group, and the DT group, 15 in each group. Rats in the sham-operation group and the model group were administered with distilled water by gastrogavage at the daily dose of 0.1 mL/kg. Rats in the DT group was administered with 0.53 g/mL DT suspension by gastrogavage at the daily dose of 0.1 mL/kg. All medication was lasted for 10 successive days. The myocardial I/R experiment was performed at 1 h after the last gastrogavage. ECG was performed before ligation and at I/R. The jugular arterial blood pressure of all rats was measured during the whole course. ST segment changes were observed at each time point of I/R. The ventricular fibrillation, the premature ventricular, the number and the duration of ventricular tachycardia within 30 min reperfusion were also observed. Activities of Na(+)-K+ ATPase and Ca2+ ATPase in the myocardium homogenate were detected as well.

RESULTSThe jugular arterial blood pressure and the heart rate were slightly lower in the DT group than in the model group, but with no statistical difference (P > 0.05). Compared with the sham-operation group, the degree of ST segment was obviously elevated in the model group at 0, 5, and 7 min (P < 0.05). It was significantly lower in the DT group than in the model group (P < 0.01). ST seg ment was more elevated at 5 min than at 0 min in the model group, but the degree of ST segment elevation was still obviously lower in the DT group than in the model group (P < 0.05). There was no statistical difference in the degree of ST segment elevation at 7 min between the two groups (P > 0.05). At 0 min when the decrement of ST segment exceeded one half the ischemia, there was no statistical difference in the degree of myocardial ischemia between the model group and the DT group (P > 0.05). Compared with the model group, the incidence of fatal and nonfatal ventricular fibrillation, the frequency and duration of ventricular tachycardia and premature ventricular beats were obviously lessened, and activities of Na(+)-K+ ATPase and Ca(2+)-ATPase increased (all P < 0.05).

CONCLUSIONSDT could significantly protect arrhythmias induced by transient I/R. Its effect might be related to lowering the degree of myocardial ischemia, and increasing ion transport channel related enzyme activities.

Animals ; Arrhythmias, Cardiac ; drug therapy ; etiology ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Male ; Myocardial Reperfusion Injury ; complications ; Rats ; Rats, Wistar

Acupuncture Therapy ; Aged ; Angina Pectoris ; therapy ; Female ; Humans ; Male ; Middle Aged

Objective: Our aim was to investigate the alteration of p16 gene in human pancreatic carcinoma. Methods: A total of 66 human pancreatic tissue specimens, comprising 51 with pancreatic carcinomas and 15 normal pancreatic tissue specimens, were examined for homozygous deletion and mutation of p16 gene by using PCR-SSCP method. Results: No mutation and deletion was detected in 15 normal pancreatic tissue samples. Of 51 pancreatic carcinoma specimens, only one was found mutation for p16 gene in PCR-SSCP assay, and the deletion of the p16 gene in 23 samples were confirmed by using PCR, with a 45% p16 gene deletion rate. Conclusion: These data suggest that p16 gene alterations may play a role in the progression of human pancreatic carcinoma.