Objective To investigate the clonal rearrangement of T cell receptor (TCR) ? gene which was of monoclonality,oligoclona-lity or clonal evolution /subclonality during the course of disease in patients with acute lymphoblastic leukemia (ALL) and its significance.Methods Between Sep. 2004 and Sep. 2007,70 patients with ALL were diagnosed in Department of Pediatrics of the First Affiliated Hospital of Guangxi Medical University,among which 51 cases were boys and 19 cases were girls.Their ages ranged from 2 years to 14 years (average age was 8.5 years).DNA samples were extracted from bone marrow cells or venous blood cells by phenol/phenol-isoamyl alcohol-chloroform/isoamyl alcohol law.DNA was amplified by polymerase chain reaction (PCR).Single strand conformation polymorphism analysis (SSCP) of silver stained technique was employed to detect PCR products.Results The amplification products of 23 cases of the 70 ALL patients were positive,in which 10 cases still had positive results in period of complete remission and had poor sensitivity to chemotherapeutic drugs,4 cases were of oligoclonality/ subclonality,and 2 cases were of clonal evolution with poor prognosis.Conclusions Detecting TCR ? gene rearrangement reflects clonal evolution of leukemia cells.The oligoclonality clonal evolution continues to exist,whose multiplication is the main reason of recurrent ALL.Detecting TCR? gene rearrangement,evaluating ALL of the patient's prognosis,the judgment of recurrence and the development of individualized treatment programs have great guiding significance,which can maximize the possibility of the sick children to make long-term disease-free survival and reduce the side effects of chemotherapy on the long-term basis.

Purpose To investigate the correlation between BRAF V600E mutation and clinicopathologic parameters in bilateral welldifferentiated thyroid carcinoma.Method Collected 80 patients with papillary thyoid carcinoma (PTC) (67 patients with classical type,8 patients with follicular variant,3 patients with oncocytic variant,2 patients with tall cell variant),5 patients with follicular carcinoma and 30 normal tissues near the tumor,BRAF V600E mutation was detected via polymerase chain reaction.Result The presence of BRAF V600E mutation was found in 52 patients with PTC (65.0％),the mutation was not in the 5 patients with ollicular carcinoma as well as in the 30 normal tissues near the tumor.The BRAF V600E mutation was positively correlated with age,capsule invasion,lymph node metastasis,tumor clinical stage.In the histological variant of PTC,the BRAF V600E mutation rate of classical type and tall cell variant is very high (70.1％,100.0％),but the BRAF V600E mutation rate of follicular variant is very low (33.3％).Conclusion The BRAF V600E mutation rates are different in different age groups.The BRAF V600E mutation was positively correlated with capsule invasion,lymph node metastasis,tumor clinical stage.The BRAF V600E mutation rate of classical type and tall cell variant is very higher than follicular variant.

To compare the clinical effect of phacoemulsification and middle incision extracapsular cataract extraction ( Ml-ECCE) . METHODS: One hundred and eighty - five eyes of phacoemulsification (137 cases) and 185 eyes of 139 cases for Ml-ECCE from January 2011 to May 2013 were involved in this study. And the ratio posterior capsular rupture during surgery, visual acuity, corneal edema, corneal astigmatism and intraocular pressure post operation were followed up.RESULTS: On 1d after surgery, uncorrected visual acuity in the group of Ml-ECCE was better than that of phacoemulsification group, while from 3d; 1 and 3mo after surgery, no significant difference was found from the above two groups. On 1d postoperation, corneal edema ratio in phacoemulsification group ( 45 eyes ) was higher than that in Ml-ECCE group(20 eyes) ( X2=11. 665, P=0. 0006 ) . No significant difference was found for the ratio of posterior capsule rupture during surgery in these two surgical technique groups(X2=0. 094,P=0. 759). On 1wk;1 and 3mo after surgery, significant difference was found for the average of surgical induced corneal astigmatism between two groups (u=6. 661, 6. 880, 4. 187, P = 0. 00, respectively ). During following up, no significant difference was found for the intraocular pressure between two groups ( u=1. 858, 0. 963, 0. 471, 1. 349, 1. 388; P= 0. 063, 0. 335, 0. 638, 0. 177, 0. 165). lntraocular pressure on 1d postoperation in phacoemulsification and Ml-ECCE groups was higher than before operation ( u = 19. 86, 19. 39, P = 0. 00, respectively). And on 1wk; 1 and 3mo postoperation, intraocular pressure in the operated eyes in both groups was lower than before operation for 2~3mmHg.CONCLUSlON: Although phacoemulsification and Ml-ECCE could both get good visual rehabilitation, with similar visual outcome, no significant effect for intraocular pressure, and no severe complications, the latter one owns the advantage that easier maneuver, quicker recovery, and cheaper instruments needed, which is suitable for the hard nuclei cataract in local hospitals. Ml-ECCE is a safe, effective and easy manipulation for local hospital for large batch of cataract surgeries.

A novel nucleic acid amplification method,termed loop-mediated isothermal amplification(LAMP),which amplifies DNA with high specificity,efficiency,and rapidity under isothermal conditions,may be a valuable tool for the rapid detection of infectious diseases.This method employs a DNA polymerase that have activity of strand displacement DNA synthesis and a set of four specially designed primers that recognize a total of six distinct sequences on the target DNA.LAMP can amplify a few copies of DNA to 109 in less than an hour.The final products are stem-loop DNA with several inverted repeats of the target and cauliflower-like structures with multiple loops.A positive reaction would be shown as a ladder-like pattern in a gel electrophoresis analysis.Because of the advantage,the LAMP method will be widely applied to research of nucleic acid,clinical diagnosis of infectious diseases and detection of genetically modified organisms etc.

Objective: To further investigate the molecular mechanism of photodynamic therapy. Methods: We used cDNA microarray technique to explore the gene expression profiles of HEPG2 cells after photodynamic therapy with hematoperphyrin monomethyl ether(HMME) in HEPG2 cells. After treated with HMME for 60 min, the HEPG2 cells were irradiated with laser, and observed by microscope with H E staining. To prepare the probes, mRNA from both control and treated cells were isolated and purified, then reversely transcribed to cDNA with the incorporation of fluorecent labeled dUTP. The probes were hybridized with a cDNA microarray representing the 1 538 genes originated from human hepatocarcinoma cells. The fluorencent signals of Cy3 and Cy5 were scanned and analyzed. Results: After laser irradiation, the HEPG2 cells showed the typical feature of apoptosis. The gene expression profiles were also changed greatly. Among the 1 538 target genes, 389(2.47%) different expression genes were detected. Most of the changed genes (nearly 80%) were down regulated. They were functionally related to cell proliferation cycle, replication, metabolism and so on. Several apoptosis associated genes were detected among those up regulated genes, encoding the key proteins involved in apoptosis signal transduction, such as CCP32,AIF,Mch2. Conclusion: The HMME photodynamic therapy can initiate the apoptosis process of HEPG2 cells, which may be regulated by mitochondial pathway.[

Aim at the two problems in the field of traditional Chinese medicine (TCM) mechanism elucidation, one is the lack of detailed biological processes information, next is the low efficient in constructing network models, we constructed an auxiliary elucidation system for the TCM mechanism and realize the automatic establishment of biological network model. This study used the Entity Grammar Systems (EGS) as the theoretical framework, integrated the data of formulae, herbs, chemical components, targets of component, biological reactions, signaling pathways and disease related proteins, established the formal models, wrote the reasoning engine, constructed the auxiliary elucidation system for the TCM mechanism elucidation. The platform provides an automatic modeling method for biological network model of TCM mechanism. It would be benefit to perform the in-depth research on TCM theory of natures and combination and provides the scientific references for R&D of TCM.
Animals ; Automation ; instrumentation ; methods ; Databases, Factual ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Gene Regulatory Networks ; Humans ; Medicine, Chinese Traditional ; Plants, Medicinal ; chemistry

Objective In order to lay the foundation for establishment of a clean grade Meriones meridianus popu-lation, and provide reference for the formulation of Meriones meridianus parasite detection standard.Methods A survey of parasite infection in Meriones meridianus was carried out, and three kinds of antiparasitic drugs, tetramethrin/per-methrin, praziquantel, and tinidazole were used in high-, medium-and low-doses for eradication of the target parasites. Results Three types of parasites were detected among the animals.The infection rate of surface parasite Myobia musculi was 82.1%(23/28), that of intestinal parasite Hymenolepis nana was 78.1% (25/32) and Tritrichomonas muris was 90.6%(29/32).Significant results of parasite eradication were achieved with the antiparasitic drugs.Medicated bath u-sing 1%tetramethrin/permethrin for 30 s every 4 days for 5 times and 1.5%tetramethrin/permethrin for 30 s every 4 days for 3 times completely eradicated the surface Myobia musculi infection.Praziquantel in a dose of 31 mg/kg by daily gastric gavage for consecutive 7 days was the optimal dosage to completely eradicate the intestinal Hymenolepis nana.Tinidazole in a dose of 42 mg/kg administered by gastric gavage daily for consecutive 7 days was the optimal dosage for eradication of in-testinal Tritrichomonas muris infection.Conclusions Three types of parasites are detected in Meriones meridianus in our laboratory, i.e.surface infection with Myobia musculi, and intestinal infection with Tritrichomonas muris and Hymenolepis nana.Tetramethrin/permethrin, praziquantel and tinidazole can be used to effectively eradicate these three types of para-sites, respectively.

Background Presbyopia is one of primary causes affecting the visual and life qualities of the agings,and its mechanism is associated with the oxidative damage of lens epithelial cells with ageing.SS31 is a mitochondria-targeted antioxidant peptide.To study the effect of SS31 on oxidative damage of lens epithelial cells has an important significance for the prevention and treatment of presbyopia.Objective This study was to investigate the effect of SS31 on in vitro oxidative damaged human lens epithelial cells.Methods Human lens epithelial cell line (HLEB-3) was cultured using DMEM with low glucose and 10％ fetal bovine serum(FBS).The cell model of oxidative damage was established by adding 200 μmol/L tea-butyl hydropeoxide (t-BHP) into DMEM for 18 hours.The cells were divided into blank control group,t-BHP model group,10 nmol/L SS31 +t-BHP group,100 nmol/L SS31 +t-BHP group,1 μmol/L SS31 +t-BHP group,10 μmol/L SS31 +t-BHP group and 100 pμmol/L t-BHP group,and then MTT assay was used to detect the survival rate of the cells and evaluate the optimal SS31 concentration for sequential study.The cells then were divided into blank control group,t-BHP model group and 1 μmol/L SS31 +t-BHP co-culture group.The change of mitochondrial membrane potential of the cells was tested by JC-1 dye and flow cytometry.Reactive oxygen species (ROS) level in the mitochondria was determined using MitoSOX staining.Results The cell survival rate in the t-BHP model group was (53.42±2.52)％,and that in the blank control group was 100％.The cell survival rate was considerably increased in various concentrations of SS31 groups,showing a significant difference among different groups (F=58.349,P＜0.01).A highest survival rate was (82.13 ±3.15) ％ in the 1 μmol/L SS31 +t-BHP co-culture group,which was statistically significant in comparison with the t-BHP model group (t =28.710,P＜0.05).JC-1 dye and flow cytometry assay showed that the ratio between red and green fluorescence intensity was 7.07 ±0.06 in the blank control group,4.46±0.14 in the t-BHP model group and 5.76±0.26 in the 1 μmol/L SS31 +tBHP co-culture group,showing significant difference among the 3 groups (F=172.332,P＜0.01).The ratios between red and green fluorescence intensity in the blank control group and 1 μmol/L SS31 +t-BHP co-culture group were higher than that in the t-BHP model (t =2.609,1.303,both at P＜0.001).ROS fluorescence cells were much more in the t-BHP model group compared with blank control group and 1 μmol/L SS31 + t-BHP co-culture group.Conclusions SS31 can protect HLEB-3 cells from oxidative stress.SS31 may serve as a potential new approach to the treatment of presbyopia and other age-related diseases of lens.

Objective To observe the effect of intermedin(IMD) on microvascular injury of renal fibrosis in unilateral ureteral obstruction (UUO) rat model.Methods Seventy-two male Wistar rats were randomly divided into two groups:the sham-operation group (n=24) underwent the left ureteral dissection,the other 48 rats were made as unilateral ureteral obstruction models and subdivided into model group(UUO,n=24) and IMD group (n=24).At the 7,14,21,28 day after the operation,6 randomly-selected rats from each of the three groups respectively were blooded by abdominal arotic and their obstructive kidneys were taken out.The renal histopathological changes were observed through HE and Masson staining,the contents of BUN,Scr and cystatin C (CysC) of the obstructive kidneys were determined,the expressions of transforming growth factor-β1 (TGF-β1),α-SMA,bone morphogenetic protein-7 (BMP-7),E-cadherin,thrombospondin 1 (TSP-1) and vascular endothelial growth factor (VEGF) were detected by RT-PCR and immunohistochemistry.Results Compared with the sham-operated group,the pathological changes of kidney in the model group showed that the degree of fibrosis was obvious,tubular interstitial damage aggravated,the levels of BUN,Scr,CysC in the model group increased (P ＜ 0.05),the mRNA expression and protein content of TGF-β1,oα-SMA,TSP-1 increased (P ＜ 0.05),while the levels of BMP-7,E-cadherin and VEGF decreased (P ＜0.05).Compared with the UUO group,renal tubular damage,interstitial fibrosis in the IMD group were lighter,the levels of BUN,Scr,CysC in the IMD group were lower (P ＜ 0.05),the mRNA expression and protein content of TGF-β1,α-SMA,TSP-1 were down-regulated (P ＜ 0.05),while the levels of BMP-7,E-cadherin and VEGF were up-regulated (P ＜ 0.05).Conclusion IMD can ameliorate the renal interstitial fibrosis,and the mechanism may be related to the fact that VEGF mediated by IMD can reduce vascular injury.

Objective To evaluate the feasibility and efficiency of transthoracic echocardiography(TTE) combined with real-time three dimensional echocardiography(RT-3DE)in guiding transcatheter closure of atrial septal defect(TCASD)without X-ray.Methods Eleven patients with atrial septal defects(ASD)underwent the procedure of TCASD guided by TTE combined with RT-3DE.The position of the catheter and transporting sheath,the location and deploying of ASD occluder(ASO)were monitored by muhisection echoscan.especially in RT-3DE.The results were compared with those guided by conventional method(TTE and X-ray).Results All the operations were held successfully.The complications of cardiac tamponade,shedding of occluder,residual shunt,atrioventricular valve regurgitation,embolism and arrhythmia were not observed.Compared with the conventional procedure of the control group,the operation time was significantly prolonged[(65.76±14.15)min versus(45.50±20.88)min,P＜0.000)].Conclusions TCASD was tested to be safe and efficient guided by TTE combined RT-3DE without X-ray,especially for children and pregnant women.It may be expected used widely in the future independently from X-ray.