OBJECTIVETo investigate the methylation status of zonula occludens-1 (ZO-1) gene promoter and its clinical significance in children with stage IV non-Hodgkin lymphoma (NHL) and to provide a basis for further etiological study and early diagnosis of this disease.

METHODSFifty-five children with a confirmed diagnosis of stage IV NHL (40 cases of T-NHL and 15 cases of B-NHL) were selected as the case group, and 20 children with diseases other than hematologic malignancies were selected as the control group. Bone marrow samples were collected from these subjects. Methylation-specific PCR (MS-PCR) was applied to evaluate the methylation status of ZO-1 gene promoter, and the integrated optical density (IOD) was determined. RT-PCR was used to measure the mRNA expression of ZO-1.

RESULTSMS-PCR showed that the methylated bands of ZO-1 gene promoter were found in 39 (70.9%) of 55 patients in the case group before treatment, while no ZO-1 gene promoter methylation was detected in the control group. With close tracking of 47 cases in the study group, consisting of 32 cases of T-NHL and 15 cases of B-NHL, the rates of ZO-1 gene promoter methylation prior to treatment were 72% and 67%, respectively, (P>0.572). The cases of T-NHL and B-NHL showed no significant changes in methylation rate in the early and middle phases of chemotherapy (P>0.05), but they showed significant changes in methylation rate in the late phase of chemotherapy (P<0.05). RT-PCR showed that the NHL cases carrying methylated ZO-1 gene had no mRNA expression of ZO-1, while all children in the control group had mRNA expression of ZO-1. There was no linear relationship between the total number of peripheral blood leukocytes and ZO-1 gene IOD (r=0.093, P=0.575); a positive correlation was found between the number of malignant cells in bone marrow and ZO-1 gene IOD (r=0.669, P<0.001).

CONCLUSIONSZO-1 gene shows a hypermethylation status in children with NHL, and the methylation level is positively correlated with the number of malignant cells in bone marrow. ZO-1 may be used as a novel molecular marker in early diagnosis, outcome assessment, prognostic evaluation, and detection of minimal residual disease.

Adolescent ; Child ; Child, Preschool ; DNA Methylation ; Female ; Humans ; Infant ; Lymphoma, Non-Hodgkin ; genetics ; Male ; Promoter Regions, Genetic ; Zonula Occludens-1 Protein ; genetics

Objective To investigate the current dyslipidemia profiles and correlation with etiological factors in early stage post-transplantation, and the impact of lipid metabolic disorder on renal function. Methods The clinical data of 1032 renal allografts from eight hospitals in Beijing between 2004 and 2008 were collected and evaluated retrospectively. Before and at the 1st, 3rd, 6th and 12th month post-transplantation, the changes in blood total cholesterol (TC), triglycerides (TG),low density lipoprotein (LDL)-cholesterol and high density lipoprotein (HDL)-cholesterol were analyzed. The difference in the blood lipid disorder at different stages stratified by time and different age group, the effects of immunosuppressive agents on blood lipid, and the impact of blood lipid disorder on the blood creatinine were studied. Results Except HDL-cholesterol, TC, LDL-cholesterol and TG levels were increased gradually at the first year, especially LDL-cholesterol and TG. The TC and LDL-choleaterol abnormalities were obviously related with age (P<0. 01 ). The effect of Tacrolimus (Tac)-based immunosuppressive regimen on the lipid metabolic disorder was less than cyclosporine (CsA). At the first year, there was no significant difference in blood creatinine between lipid-lowering treatment and non-lipid-lowering treatment (P>0. 05). For the recipients not subject to lipid-lowering treatment and their TG level higher than the normal at the first month after operation, the creatinine level at the first year was significantly higher than in those with normal TG level (P< 0. 05). Conclusion The lipid metabolic disorder following renal transplantation is a common complication after the first transplant year, and was related with age and immunosuppressive agent regimen. Tac-based immunosuppressive regimen has little effects on the blood lipid metabolism.

The content of gamma-aminobutyric acid in the aerial part of Panax notoginseng in different productive area was determined by using high performance liquid chromatography. HPLC analysis was made on a C18 (4.6 mm x 250 mm,5 microm) with acetonitrile and water containing 4.1 g x L(-1) sodium acetate as mobile phase. The detection wavelength was 254 nm. The HPLC method showed good linearity within the range of 0.01 - 1.03 g x L(-1). The average recovery of GABA in the stems and leaves of P. notoginseng and the flowers of P. notoginseng was 101.7% (RSD 1.1%, n = 3) and 97.3% (RSD 0.38%, n = 3), respectively. The contents of GABA in the samples of different productive areas were not significant different, and the average contents of GABA in the stems and leaves of P. notoginseng and the flowers of P. notoginseng were 0.49% and 0.53%. This method was simple and reliable, and it was suitable for the determination of gamma-aminobutyric acid in the aerial part of P. notoginseng.
Chromatography, High Pressure Liquid ; methods ; Flowers ; chemistry ; Panax notoginseng ; chemistry ; Plant Extracts ; analysis ; Plant Leaves ; chemistry ; gamma-Aminobutyric Acid ; analysis

Objective To summarize and analyze the epidemiological and clinical characteristics of COVID-19 Omicron variant cases in makeshift hospital, and the influence of age, sex and vaccination status on the disease duration, so as to provide reference for the prevention and control of the COVID-19 epidemic. Methods The epidemiological and clinical characteristics of COVID-19 cases admitted to makeshift hospital of National Convention and Exhibition Center (Shanghai) from April 9 to May 31, 2022 were retrospectively described and analyzed, and further cohort analysis was conducted to determine the influence of age, sex and vaccination status on the disease duration of COVID-19 cases in the author's branch hospital. Results Among the 174 466 COVID-19 cases in makeshift hospital, most of them were male, accounting for 59.38%. The infected cases were mainly young and middle-aged people aged 18-59 years old, accounting for 83.50%, followed by 12.30% of the elderly group over 60 years old; the average hospital stay was 7.40 days; the proportion of patients with fever was less than 27.79%; 15.37% (26 817/174 466) of the patients complicated with underlying diseases, and the top three were hypertension, diabetes and coronary heart disease. The proportion of people who received COVID-19 vaccine accounted for 79.56% (13 799/17 956), of which the highest proportion of three doses was 44.09%. The disease duration of 17 956 COVID-19 cases in the author's branch of makeshift hospital was 10.18 (7.34, 13.05) days. The disease duration in the elderly group was the longest with 11.34 (8.35, 14.37) days, followed by 11.17 (9.07, 14.33) days in the preschool group, 10.37 (8.14, 13.34)· days in the middle-aged group, 10.07 (7.37, 12.37) days in the school-age group, and 9.34 (7.05, 12.16) days in the young group. There was significant difference in the overall distribution of disease duration among the five groups (H=550.479 P<0.01). The disease duration in each age group basically showed a V-shaped distribution. The disease duration was 10.27 (7.34, 12.57) days in males and 10.10 (7.25, 13.09) days in females, and there was no significant difference (Z=-1.505 P>0.05). The disease duration of vaccinated patients was 10.24 (7.35, 13.05) days, and that of unvaccinated patients was 9.47 (7.09, 12.47) days. There was significant difference between the two groups (Z=-4.338 P<0.01). Conclusions COVID-19 Omicron variant cases have a high proportion of males, mainly young and middle-aged, and the proportion of fever patients is less than 30%. The disease duration is significantly lower than that of the original strain in Wuhan, and shows "V" distribution with each age group. Sex had no effect on the disease duration. COVID-19 vaccination did not have a clinical effect on the disease duration.

Objective To investigate the relationship between watching television time and impaired glucose regulation (IGR) , type 2 diabetes mellitus in Chongqing City .Methods Population-based cross-sectional study was conducted to investigated the local permanent staff(lived in Chongqing more than 5 years) who were 40 years old or elder in Chongqing City .Results The overall prevalence rate of IGR and T 2DM was 6 .3% ,5 .6% respectively .The average weekly watching TV time of the samples was (12 .3 ± 10 .1) h .After adjusting for possible confounding factors ,the prevalence rate of IGR and T2DM in patients watching TV time >14 h per week was significantly higher than those watching TV time ≤ 7 h per week(Adjust .OR = 1 .528 ,95% CI = 1 .034 - 2 .121 ;OR = 1 .482 ,95% CI = 1 .133 - 2 .047 ,respectively ) .Conclusion Siting and watching TV time were positively correlated with the risk of IGR ,T2DM .So ,we should actively encourage and promote healthy lifestyles to reduce siting and watching TV time .

Moderate drought stress has been found to promote the accumulation of active ingredients in Glycyrrhiza uralensis root and hence improve the medicinal quality. In this study, the transcriptomes of 6-month-old moderate drought stressed and control G. uralensis root (the relative water content in soil was 40%-45% and 70%-75%, respectively) were sequenced using Illumina HiSeq 2000. A total of 80,490 490 and 82 588 278 clean reads, 94,828 and 305,100 unigenes with N50 sequence of 1,007 and 1,125 nt were obtained in drought treated and control transcriptome, respectively. Differentially expressed genes analysis revealed that the genes of some cell wall enzymes such as β-xylosidase, legumain and GDP-L-fucose synthase were down-regulated indicating that moderate drought stress might inhibit the primary cell wall degradation and programmed cell death in root cells. The genes of some key enzymes involved in terpenoid and flavonoid biosynthesis were up-regulated by moderate drought stress might be the reason for the enhancement for the active ingredients accumulation in G. uralensis root. The promotion of the biosynthesis and signal transduction of auxin, ethylene and cytokinins by moderate drought stress might enhance the root formation and cell proliferation. The promotion of the biosynthesis and signal transduction of abscisic acid and jasmonic acid by moderate drought stress might enhance the drought stress tolerance in G. uralensis. The inhibition of the biosynthesis and signal transduction of gibberellin and brassinolide by moderate drought stress might retard the shoot growth in G. uralensis.
Droughts ; Gene Expression Regulation, Plant ; Glycyrrhiza uralensis ; genetics ; Plant Roots ; Sequence Analysis, DNA ; Stress, Physiological ; Transcriptome

Objective To investigate the possible role of interstitial cell of Cajal (ICE)in the pathogenesis of diabetic gastroparesis and the protective effect of"Xiaopi prescription (消痞方)".Methods Fifty healthy male Spregue-Dawley (SD) rats were randomly divided into 5 groups (each n=10):normal, model,and 3 Xiaopi prescription groups:low,middle and high dosages.Diabetes was induced by intravenous injection of alloxan,and equal amount of normal saline was intravenously injected in the normal group.Gastric lavage method was used to administer the traditional Chinese medicine decoction of Xiaopi prescription in corresponding amount (5,10 and 20 g?kg~(-1)?d~(-1)) in respective low,middle and high dosage groups.In the normal control group and diabetic model group,only equal amount of normal saline was administered into the stomach.Gastric emptying rate was measured by method of nutritious semisolid paste;c-kit positive cells of ICC were quantitatively measured with immunohistochemistry assay and computer image analysis system;c-kit mRNA positive cells were quantitatively measured with in situ hybridization and computer image analysis system.Results①Gastric emptying rate:The rate was significantly lower in the model group than that in the normal control group (P0.05),but higher than those in the model group and the low dosage group (all P0.05).②c-kit immmunohistochemistry:c-kit positive cell presented yellow in color,and its membrane was stained yellow,this kind of cells primarily were distributed around the neural plexus in the inter-space between the circular and longitudinal muscular fibers, and around the ganglionic cells forming"sheath-like"structure.The results of numbers of c-kit positive cells in the various groups:the number of the cells in the model group was significantly lower than that in the normal group (P0.05),but the numbers in the former two dosage groups were obviously higher than those in the model group (all P0.05),being significantly lower than that in the normal group,middle and high dosage groups (all P0.05),but obviously higher than those in the model group (all P0.05),being significantly lower than that in the normal,middle and high dosage groups (all P

Objective To observe the morphologic changes of peripheral blood lupus cell in patients with systemic lupus erythematosus and investigate its relationship with disease activity in systemic lupus ery- thematosus.Methods Modified classical blood clotting method to observe the morphological changes of pe- ripheral blood lupus cells in 80 cases with systemic lupus erythematosus.Fifty cases were in active stage,and 30 cases were in stable stage.Comparison of serum autoantibody,complement and SLEDAI were also carried out.Results There was significant association between lupus cells in special morphous and autoantibody, such as anti-double-stranded DNA antibody,anti-nucleosome antibodies,complement C3、C4 and SLEDAI(r= 0.588,P=0.056:r=0.759,P=0.135;r=-0.648,P=0.058;r=-0.589,P=0.057,r=0.686,P

OBJECTIVETo investigate the feasibility of life span extension of transformed human embryonic tendon cells (THETC) by reconstitution of the telomerase activity.

METHODSTHETC were transfected by pGRN145 plasmid containing the human telomerase reverse transcriptase (hTERT) cNDA in vitro by molecular cloning technique. The biological characteristics of transfected cells were detected and compared by morphological observation, plate cloning efficiency, soft agar culture, growth curve of cells cultured in different conditions, immunohistochemistry, telomerase activity assay by telomeric repeat amplification protocol (TRAP).

RESULTSThe THETC transfected by pGRN145 plasmid (telT) could express the telomerase activity with extension of life span. The telT maintained the original characteristics of temperature-dependant and serum-dependant, as well as secretion of type I collagen normally and without tendency of malignant transformation.

CONCLUSIONSThe life span of THETC can be prolonged by reconstitution of telomerase activity, which provides the novel experimental methods to establish the standard cells line.

Cell Line ; Cell Survival ; Embryo, Mammalian ; Humans ; Plasmids ; genetics ; RNA-Directed DNA Polymerase ; Telomerase ; genetics ; metabolism ; Tendons ; cytology ; enzymology ; Transfection

OBJECTIVETo explore the mechanism of warming-needle therapy in treatment of knee osteoarthritis of deficiency-cold syndrome.

METHODSEight cases of knee osteoarthritis of deficiency-cold syndrome were selected and treated with warming-needle therapy at Guanyuan (CV 4), Qihai (CV 6) , Zosanli (ST 36), etc.. The gene expression profiles before and after treatment in 4 cases who showed better therapeutic effect were compared. Taking ratio < 0.5 or ratio > 2.0 as differentially expression gene and obtaining differentially expression pathway (P < 0.5, n>3) by http://www. DAVID 2006.

RESULTSTwo cases were clinically cured, 4 cases were markedly effective, 1 case was effective and 1 case was ineffective. With help of the microarray, 449 differentially expression genes, and 10 differentially expression pathways were obtained including 2 energy metabolism pathways (oxidative phosphorylation, ATP synthetase), 4 cell signal transduction pathways (insulin signal pathway, Toll-like receptor signal pathway, JAK-STAT signal pathway, and MAPK signal pathway) and cell apoptosis pathway.

CONCLUSIONWarming-needle therapy is an effective therapy for knee osteoarthritis with deficiency-cold syndrome , which is possibly involved in the control and regulation of many gene expression by various signal transduction pathways.

Acupuncture Therapy ; methods ; Aged ; Female ; Gene Expression Profiling ; Gene Expression Regulation ; Humans ; Janus Kinases ; genetics ; MAP Kinase Signaling System ; Male ; Middle Aged ; Myeloid Differentiation Factor 88 ; genetics ; Needles ; Osteoarthritis, Knee ; metabolism ; therapy ; STAT Transcription Factors ; genetics ; Yang Deficiency ; metabolism ; therapy