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.

Objective To find out the distribution characteristics of drinking water with high arsenic in Dali Prefecture, Yunnan. Methods General investigation plus sampling survey was adopted in the city of Dali and 11 counties. The arsenic content in water was tested by half-quantitative fast reagent-box method. The water samples exceeding the standard(≥0.03 mg/L) were re-tested by silver diethyldithiocarbamate eolorimetric method or mercury-atomic fluorescence spectrometric method. Population and children exposed by high-arsenic were statistically analyzed. Results Arsenic content in 15 180 samples from 2639 villages are screened, of which 14 976 samples were less than 0.01 mg/L, reaching 98.66% (14 976/15 180); 110 samples was no less than 0.05 mg/L, only accounting for 0.72%(110/15 180). Water sources with excessive arsenic was found in 29 villages, in a percentage of 1.1% of all covered villages(29/2639). The samples were constituted of 10 399 portions of well water(well was less than 10 m deep), 3903 from spring, 93 from river water, 69 from hot spring water, 26 from reservoir water and 690 from surface water. And for the samples which arsenic content were ≥0.05 mg/L, 89 samples(0.86%, 89/10 399) were from well water, 15 from spring water(0.38%, 15/3903) and 6 from spring water(8.70%, 6/69). A total of 1 561 553 individuals were investigated, in a percentage of 67.83%(1 561 553/2 302 156) of the whole population, among those 420 513 were children, rating 26.93% of the investigated population(420 513/1 561 553); 27 865 were exposed to arsenic, accounting for 1.78% of the investigated population 27 865/1 561 553; 8993 children were exposed, rating 2.14% of the investigated population(8993/420 513). Conclusions There exists high-arsenic water resources in Dali Prefecture, Yunan, so the local inhabitants are in the danger of high-arsenic exposure. Urgent attention shall be paid for the endemic arsenic including investigation, prevention and control.

A procedure for the preparation of PCR template from Saccharomyces cerevisiae using boiling method is described,and arg-13 gene from low copy ARSCEN plasmid and ymc1 gene from genomic DNA are amplified with high efficiency respectively.

The inhibitory ratio (1%) of artificial musk on cyclooxygenase-2 (COX-2) was determined by enzyme immunoassay (EIA). The dose-effect relationship between concentrations of artificial musk and 1% was established. It was found that artificial musk had obvious inhibitory action on COX-2. The concentration for 50% of maximum inhibitory effect (IC50) was about 2.26 mg x mL(-1). There was a good relationship between the logarithm concentrations of artificial musk and 1% when the concentrations of artificial musk ranged from 0.31-20.0 mg x mL(-1). The results indicated that this EIA method could be applied to evaluate the anti-inflammatory activity of artificial musk quickly, conveniently, sensitively and exactly. This paper provided a novel method and foundational research for the bioassay of artificial musk.

OBJECTIVETo investigate the abnormal change of immune function in patients with Pi-Qi deficiency Syndrome, and to explore the genomic mechanism of its genesis by cDNA chip techniques.

METHODSThe cross probe was made by extracting and microamplifying the total RNA and mRNA of peripheral white blood cells (WBC) in healthy subjects and patients with chronic gastritis and ulcerative colitis, which were labeled by Cy3 and Cy5 respectively. Then equal quantity of the two labeled probes were mixed and hybridized with cDNA chip, fluorescent signal of the chips were scanned with scanner. Data obtained were analyzed for comparing the difference of the expressive levels of immune associated genome in peripheral WBC in healthy subjects with those in patients.

RESULTSExpressions of CD9, CD164, PF4 and RARB gene in WBC of patients, both gastritis and colitis, were down-regulated while those of IGKC, DEFA1 and GNLY were up-regulated.

CONCLUSIONThe genesis of Pi-Qi deficiency syndrome has its immune associated genomic basis, and the immune functions are disordered in patients with that syndrome.

Adult ; Colitis, Ulcerative ; genetics ; immunology ; Diagnosis, Differential ; Gastritis ; genetics ; immunology ; Genome ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; Qi ; Splenic Diseases ; genetics ; immunology ; Yang Deficiency ; genetics ; immunology

Objective To investigate the effect of HbA1c meeting the standard or not on microalbuminuria,blood lipids and liver enzymes in patients with type 2 diabetes.Methods A retrospective analysis was performed on 457 subjects who had type 2 diabetes.They were divided into substandard group and standard group according to HbA1c result.The general information and relevant laboratory indicators of patients were.collected and compared between two groups.Results The microalbuminuria,serum triglyceride and liver enzymes (glutamyl transpeptidase,alkaline phosphatase,aspertate aminotransferase) were significantly different between two groups [ (189.8 ± 235.3) mg/dl vs (38.9 ± 85.5) mg/dl,(2.64 ± 2.99) mmol/L vs (2.02 ± 1.50)mmol/L,(41.7 ±52.9)U/L vs (29.7 ±24.9)U/L,(83.6 ±28.6) U/L vs (74.3 ±25.8)U/L,(26.7 ±19.1)U/L vs (22.0 ±10.5) U/L,P ＜0.05].HbA1c level was positively correlated with microalbuminuria,glutamyl transpeptidase and alkaline phosphatase (r =0.209,0.115,0.11,P ＜0.01).The microalbuminuria was an independent risk factor of affecting HbA1c to reach the standard (OR = 1.009,P ＜0.05).Conclusions HbA1c meeting the standard or not can influence many factors except blood glucose.

ObjectiveTo investigate the correlation between neutrophil to lymphocyte ratio (NLR) and cerebral microbleeds (CMBs) in patients with acute ischemic stroke.MethodsThe consecutive inpatients with acute ischemic stroke were prospectively enrolled.Gradient echo-T2*-weighted imaging was used to evaluate CMBs and their quantity.Univariate analysis was used to compare the baseline data between the CMB group and the non-CMB group.Multivariable logistic regression analysis was used to identify the independent correlation between NLR and CMBs.ResultsA total of 218 patients with acute ischemic stroke were prospectively enrolled, including 66 (30.3%) with CMBs.The age (64.7±6.6 years vs.66.9±8.6 years;t=2.052, P=0.041), high sensitive C-reactive protein (7.0[2.3-13.9] mg/L vs.8.9[4.0-28.1] mg/L;Z=2.008, P=0.045) and NLR (1.9[1.4-2.9] vs.2.3[1.7-3.6];Z=2.071, P=0.038) in the non-CMB group were significantly lower than those of the CMB group.Multivariate logistic regression analysis showed that NLR (odds ratio 1.276, 95% confidence interval 1.008-1.670;P=0.045) and age (odds ratio 1.044, 95% confidence interval 1.002-1.087;P=0.040) were the independent risk factor for CMBs.Spearman correlation analysis showed that NLR was significantly positively correlated with the severity of CMBs (r=0.210, P=0.007).ConclusionsIn patients with acute ischemic stroke, NLR was associated with CMBs and their severity, suggesting that inflammatory reaction might be involved in the occurrence of CMBs.

Background Oxidative damage may cause the functional dysfunction and death of retinal vascular endothelial cells(RVECs),and further leads to the development of retinal vascular diseases.Fufang xueshuantong has a therapeutic effect on retinal vascular diseases,but little is known about its molecular mechanism.Objective The goal of this study was to investigate the protective effects and mechanism of fufang xueshuantong on injury of human RVECs induced by tert-butyl hydroperoxide(t-BHP).Methods Human RVECs were isolated from healthy donor eyes and primarily cultured and then identified by flow cytometry.The third to fifth generations of cells were used in this experiments.The fufang xueshuantong solution of 0.0625,0.1250,0.2500,0.5000 and 1.0000 g/L were added in the cuhure plate with 5 × 104/L cells respectively in the experimental groups,and t-BHP of 75,100,200 and 300 μ.mol/L were added in the model control groups.MTT was used to detect the A490and survival rate of RVECs.The apoptotic rate and death rate of the cells were evaluated by double staining of Annexin V-FITC/PI.Morphology of human RVECs were examined using invert microscopy and Hoechst33258 staining.The expressions of nitro tyrosine (a marker of oxidative damage of protein)and 8-OHdG(a marker of oxidative damage of DNA)in human RVECs were assessed by the immunofluorescence staining.Western blot was used to detect the expressions of nuclear factorkappa B(NF-KB),p53,bcl-2 and bax after 6,12,24 hours t-BHP action.This study was approved by the Ethic Committee of Zhongshan Ophthalmic Center.Results No significant difference was found in A490value among the normal control group,0.0625,0.1250,0.2500,0.5000 and 1.0000 g/L fufang xueshuantong groups(F =1.989,P＞0.05).The survival rates of the cells were lower in 75,100,200 and 300 μmol/L t-BHP groups compared with corresponding fufang xueshuantong groups(t =14.57,13.82,21.51,32.64,P＜ 0.01).The percentages of normal cells were evidently lower in 75,100,200 and 300 μmol/L t-BHP compared with corresponding fufang xueshuantong groups(t=14.908,5.495,17.165,26.330,P＜0.01).The numbers of deformation and death of the human RVECs increased as the elevated concentration of t-BHP,but those in fufang xueshuantong groups were less than the t-BHP groups under the invert microscopy.Compared with t-BHP groups,the expressions of nitro tyrosine,8-OHdG,NF-KB,p53 and bax were lower but the expression of bcl-2 was higher in human RVECs with the statistically significant differences(P＜0.05).Fufang xueshuantong at the concentration of 0.2500 g/L showed maximally protective effect on human RVECs.Conclusions Fufang xueshuantong protects human RVECs against the t-BHP-induced injury through downregulating the expression of NF-kB,p53,bax and up-regulating the express of the bcl-2 protein.

Objective To explore the effects of modified tubo-uterine implantations performed on women with proximal tubal occlusive infertility after femal sterilization with mucilago phenol.Methods Two hundred and eight infertile women who were admitted to the Second Affiliated Hospital of Sun Yat-sen University between 1986 and 2004 were included.They all accepted modified tubo-uterine implantation after occlusion of fallopian tubes with mucilago phenol.Results It was found that the occlusions were all located in the interstitial portion or isthmic portion of the fallopian tubes.Different degrees of pelvic adhesions were found in 65 cases.Fifty-seven cases were slightly adhesive,seven cases were of moderate degree and one case was severe.One hundred and ninety-nine cases were followed up after operations(95.7%).One hundred and ninety-three women accepted hydrotubation in the following month just after the operation and 185 women were found to be unobstructed(95.8%).One hundred and forty-three women became pregnant, the pregnant rate being 71.9%(143/199).One hundred and twenty-five women had term deliveries (87.4%),three women were in early pregnancy and two in midtrimester pregnancy.Eleven women had spontaneous abortion(7.7%).Two women had tubal pregnancy(1.0%).None of the 199 cases had any signs of endometriosis.Conelusions Modified tubo-uterine implantations are quite effective for proximal tubal occlusive infertility.It may be a favorable method for such kind of tubal occlusions.

OBJECTIVEConditioned taste preference (CTP) is a taste learning reflex by which an animal learns to prefer a substance which tastes not well and has been studied with much interest in recent years. However, the neural substrates of CTP are less known. This study aimed to determine the possible neural path- ways of CTP and whether serum leptin level and the leptin receptor (OB-Rb) in the hind brain are involved following CTP formation.

METHODSWe established CTP of quinine in rats with a 2-bottle preference test. The serum leptin concentrations were detected, the expression of c-fos in the rat brain was tested to determine the nuclei in relation with establishment of CTR Finally, the OB-Rb mRNA expression was examined by RT-qPCR assay in parabrachial nucleus (PBN) and the nucleus of the solitary tract (NST) of the hind brain.

RESULTSCompared with control group, the level of serum leptin was higher in the CTP group (4.58 ± 0.52 vs 1.67 ± 0.25 µg/L, P < 0.01); increased c-fos positive cells were found in the anterior hypothalamus (AH, 221.75 ± 4.96 vs. 178.50 ± 6.63 cells/mm², P < 0.05), the basal lateral amygdala (BLA, 70.75 ± 6.17 vs 56.50 ± 3.62 cells/ mm², P < 0.05) and the nucleus of the solitary tract (NST, 41.25 ± 1.32 vs 32.50 ± 1.02 cells/mm², P < 0.05). But in ventromedial nucleus of the hypothalamus (VMH, 20.75 ± 2.73 vs 38.5 ± 1.54 per 1 mm², P < 005), PBN (21.50 ± 2.24 vs 36.25 ± 1.49 cells/mm², P < 0.05) and the central nucleus of the amygdala (CeA, 22.25 ± 1.53 vs 35.50 ± 2.11 cells/mm², P < 0.05), the number of c-fos positive cells was decreased in the CTP group. In addition, we found OB-Rb mRNA expression in PBN of CTP group rats was higher than that of control group (0.95 ± 0.055 vs 0.57 ± 0.034, P < 0.05), while there was no significant difference of OB-Rb mRNA expression in NST between the two groups.

CONCLUSIONNuclei AH, BLA, NST, VMH, PBN and CeA participate in the formation of CTP. Leptin and its receptor in PBN may be involved in the formation and maintenance of CTP.

Animals ; Conditioning (Psychology) ; Leptin ; blood ; Rats ; Receptors, Leptin ; physiology ; Rhombencephalon ; physiology ; Taste ; physiology