A total of 118 patients with type 2 diabetes mellitus were divided into acarbose treatment group ( A group,n =58 ) and no acarbose treatment group ( B group,n =60),and 57 healthy subjects were used as control group (C group).The quantification of fecal bifidobacteria and enterococcus faecalis in these subjects was made by realtime PCR.The results showed that fecal bifidobacteria contents in A and B groups were lower and enterococcus faecalis contents were higher compared with C group.After four weeks of intervention,fecal bifidobacteria contents in A and B groups increased ( P＜0.01 ),especially in A group,while enterococcus faecalis contents decreased ( P＜0.05 )compared with the baseline.Univariate correlation analysis showed that bifidobacteria content was negatively associated with lipopolysaccharides(LPS),advanced glycation index,high sensitive C reactive protein ( hs-CRP),and body mass index ( BMI ) at baseline ( P＜0.05 or P＜0.01 ).The enterococcus faecalis content was positively associated with levels of monocyte chemoattractant protein-1,LPS,tumor necrosis factor-α,hs-CRP,plasminogen activator inhibitor-1,BMI,and HbA1c (P ＜0.01 ).After four weeks of intervention,the above associations disappeared.Stepwise multivariate regression showed that basal BMI,HbA1c,and age contributed to the increase in the number of enterococcus faecalis,and BMI negatively contributed to the decrease in number of bifidobacteria.

Objective To investigate the effects of subclinical hypothyroidism (SCH) on blood glucose and metabolic parameters in type 2 diabetes (T2DM) . Methods A total of 973 T2DM patients were recruited, and the clinical data, biochemical and thyroid function parameters were measured. One hundred and forty eight T2DM patients were diagnosed as SCH (SCH group) and 152 euthyroid patients were selected as control group (NSCH group) . Results (1) The prevalence of SCH in type 2 diabetes was 15.3% (148/973) . (2) Compared with the NSCH group, the prevalence of overweight/obesity was significantly higher in SCH group (54.1%vs 35.5%,<0.01) as well as hypertension (64.9%vs 52%, <0.05) . There was no significant difference in HbA1c control and prevalence of dyslipidemia and hyperuricemia between two groups. Conclusion SCH may have some adverse effects on blood pressure and BMI in T2DM.

An HPLC fingerprint of pomegranate peel was established. Using chromatographic conditions, we compared the chemical composition of pomegranate peel, inside and seeds, and simultaneously determined the contents of gallic acid and ellagic acid. By comparison, we found that there were no significant differences between pomegranate peel and inside, but there was a big difference between pomegranate seeds and another two. The contents of gallic acid and ellagic acid of pomegranate peel respectively were 0.33%, 0.59%, while in pomegranate inside the result respectively were 0.52%, 0.38%. Content of ellagic acid from pomegranate seeds was only 0.01%. By study, we thought that when pomegranate peel was processed, pomegranate seeds should be removed, while pomegranate inside could be retained on the premise of full drying.
Ellagic Acid ; analysis ; Gallic Acid ; analysis ; Punicaceae ; chemistry ; Seeds ; chemistry

OBJECTIVESTo investigate whether antisense human telomerase reverse transcriptase (hTERT) could inhibit the activity of telomerase and the proliferation of K562 cells.

METHODSThe antisense plasmid was constructed by reverse insertion of hTERT PCR product into plasmid pLNCX-neo. Then the constructed plasmid was introduced into K562 cells by liposomes-mediated DNA transfection. The inhibition effects of telomerase on the proliferation of K562 cells were analyzed by MTT and colony formation assay, the telomerase activity of K562 cells by TRAP-PCR ELISA methods.

RESULTSThe growth rate of antisense hTERT transfected K562 cells was significantly lower than those of the controls, and the colony formation capacity of the transfected cells decreased significantly (P < 0.01), the colony number is (100.33 +/- 7.57)/10(3) cells, (92.67 +/- 5.86)/10(3) cells and (50.33 +/- 6.11)/10(3) cells for control K562 cells, K562 neo cells and antisense hTERT transfected HL60 cells, respectively. The telomerase activity of antisense hTERT transfected K562 cells was significantly inhibited.

CONCLUSIONThe expression of an antisense sequence to the mRNA sequence of telomerase protein subunit can inhibit the activity of telomerase, slow the cell growth and inhibit the capacity of colony formation of K562 cells.

Cell Division ; drug effects ; Humans ; K562 Cells ; Plasmids ; genetics ; RNA, Antisense ; genetics ; pharmacology ; RNA, Messenger ; genetics ; Telomerase ; drug effects ; genetics ; metabolism ; Transfection

Objective To evaluate the surgical treatment for renal cell carcinoma with inferior vena cava tumor thrombus and the clinical significance of multidisciplinary treatment. Methods Two cases of renal cell carcinoma with inferior vena cava thrombus diagnosed by Doppler ultrasonography and CT were included in this retrospective analysis. The tumor thrombus was in level Ⅱ in one case and in level Ⅳ in the other. Coagulation test and complete blood count were done again before surgery. Human albumin, fibrinogen, prothrombin complex, plasma, platelet, UW and irrigating solution were prepared before the operation.Under general anesthesia, surgery was performed using abdomen inverted Y shaped incision. Right radical nephrectomy was finished by the urological surgeon; the vena cava was completely dissected from the renal vein level to the secondary porta of the liver by the hepatobiliary surgeon, the vena cava and the surrounding branch vein were blocked in the upper and lower vena cava tumor thrombus; tumor thrombus was removed completely by the vascular surgeon. In one case (patient with level Ⅳ thrombus ) where the tumour thrombus invaded the wall of the vena cava, the thrombus was found to be extending to the cavo-atrial junction but not into the right atrium. The left femoral venous-right atrial bypass was established, the cardiopulmonary bypass lasted for 241 mia, and the aorta was blocked for 18 min. Salvage autotransfusion was used during surgery, and the hepatic vein of the secondary liver porta was anastomosed to artificial vascular graft.The data for surgical indication, operation time, operative blood loss and postoperative hospital stay were analyzed. Results Right radical nephrectomy and inferior vena cava thrombectomy were performed successfully, and the two patients were discharged on the 15th and 27th day after surgery, respectively. The two patients were followed up for 1 and 16 months after surgery, respectively, and both survived without local recurrence and distant metastasis. Conclusion Radical nephrectomy and inferior vena cava thrombectomy is the preferred method for patients without metastasis, and multidisciplinary cooperation could shorten the operation time, reduce the tumor recurrence and increase the survival rate of patients.

OBJECTIVETo investigate the absolute bioavailability of caffeic acid in rats and its intestinal absorption properties.

METHODThe absolute bioavailability (Fabs) of caffeic acid was obtained after iv (2 mg x kg(-1)) or ig (10 mg x kg(-1)) administration to rats. The intestinal absorption of caffeic acid was explored by the recirculating vascularly perfused rat intestinal preparation. Caco-2 cell model was applied to measure the permeability of caffeic acid from apical to basolateral said (A-B) and from basolateral to apical said (B-A).

RESULTA two-compartment pharmacokinetic model was best to describe the pharmacokinetics of caffeic acid following iv or ig administration. The Fabs of caffeic acid was 14. 7% , and its intestinal absorption was 12.4%. The values of Papp A-->B and Papp B-->A of caffeic acid were retained stable while its concentration was changed. The efflux ratio values in this study surveyed were above 2.0, and suggesting caffeic acid was active transport.

CONCLUSIONCaffeic acid was shown to have poor permeability across the Caco-2 cells, low intestinal absorption and low oral bioavailability in rats.

Animals ; Biological Availability ; Caco-2 Cells ; Caffeic Acids ; metabolism ; pharmacokinetics ; Humans ; Intestinal Absorption ; Male ; Rats ; Rats, Sprague-Dawley

Yunnan sudden death syndrome (YSDS) is an abruptly fatal disease of unknown etiology, found mostly in central or northwestern mountain area (with altitude between 1,815 and 2,225 meters) of Yunnan province from June to September every year. It occurs mostly in young female adults, with high incidences in Lisu, Yi and Miao ethnics and high familial aggregation. The clinical manifestation of YSDS is changeful and the pathological characteristic is lack of specificity. The pathogenesis may be attributed to several factors including poor hygiene and lower socioeconomic conditions, lack of Selenium or Chromium, infection of Coxsackie B virus, mushroom consumption and special geological conditions. This article reviews the epidemiologic features, clinical manifestations, pathological features, etiology and hypothesis in order to provide clues for the research of YSDS.
Adult ; China ; Death, Sudden/pathology* ; Female ; Humans ; Syndrome

Objective To evaluate the effect of acarbose on the circulating concentration of inflammatory factors in patients with type 2 diabetes mellitus(T2DM). Methods Total 118 patients with T2DM who did not take acarbose before enrollment within 4 weeks were recruited by a randomizing formula into 2 groups ( group A and B). 57 healthy subjects were included into group C as control. After excluding those of inadequate samples, 57 patients with T2DM were enrolled into group A in which acarbose was prescribed 50 mg three times a day, 59 patients with T2DM were enrolled into group B in which acarbose was not given and other hypoglycemic approaches were similar to group A. Serum samples at the time of enrollment and at the end of 4 weeks intervention were collected and stored in refrigerator at -80℃ until analysis. Analysis of biochemical indexes was performed in central lab of the institution,inflammatory factors were determined with commercial ELISA kits. Results (1) The metabolic indexes were significantly decreased after intervention in two diabetic groups. (2) The baseline levels of monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-α( TNF-α), prothrombin activator inhibitor-1 (PAI-1), lipopolysaccharide (LPS), high sensitive C reactive protein ( hs-CRP), and advanced glycation index (AGI) in diabetic patients were significantly higher than in group C MCP-1 [( 463.4± 187.1 vs 267.1 ± 158.3 ) pg/ml, TNF-α( 12.07 ± 19.59 vs 4.18 ±3.03 ) pg/ml, PAI-1 ( 2.47 ± 1.87 vs 1.38 ± 2.37 )ng/ ml, LPS ( 130.6 ± 128.5 vs 29.39 ± 17.93 ) pg/ml, hsCRP(4.25 ±2.29 vs 2.11 ± 1.07 ) μg/ml, AGI (3.78 ± 2.61 vs 0. 74 ± 0. 15 ) AU, all P ＜ 0. 05]. (3) Repeated measurement ANOVA analysis showed that after four weeks of intervention, MCP-1 [F( 1,106 ) = 19. 830, P＜0.001],LPS[F(1,106)=7.815, P＜0.01], PAI-1 [F(1,106)= 7.792, P＜0.01], TNF-α[F(1,106=24. 656, P=0.001 )] ,AGI[F( 1,106)= 12. 971 ,P=0. 01] decreased significantly in group A than in group B. Although hsCRP decreased in both group A and group B, but the trend was not different [F( 1,102 )= 0. 915, P = 0. 342].Conclusion The levels of inflammatory factors were elevated in patients with type 2 diabetes mellitus, which could be mostly reduced by acarbose.

OBJECTIVETo construct a single plasmid vector mediating doxycycline-inducible recombined human insulin gene expression in myotube cell line.

METHODSAn expression cassette of rtTAnls driven by promoter of human cytomegalovirus and a furin-cuttable recombined human insulin expression cassette driven by a reverse poly-tetO DNA motif were cloned into a single plasmid vector (prTR-tetO-mINS). The prTR-tetO-mINS and pLNCX were co-transfected into a myotube cell line (C2C12) and pLNCX vector were used as a control. After selection with G418, the transfected cells were induced with doxycycline at concentrations of 0, 2, and 10 microg/mL. RT-PCR was used to determine expression levels of recombinant insulin mRNA at the 5th day. Insulin production in cell cultures medium (at different incubation time) and cell extracts (at the 7th day) were analyzed with human pro/insulin RIA kits.

RESULTSImmune reactive insulin (IRI) level in cell medium was found increased at 24 hours of doxycycline incubation, and still increased at the 5th day. After withdrawn of doxycycline, IRI decreased sharply and was at baseline three days later. IRI and human insulin mRNA levels were positively related to different levels of doxycycline. A 25-fold increase in IRI was found against background expression at the 7th day.

CONCLUSIONHuman insulin expression can be successfully regulated by doxycycline and the background was very low. This single tet-on insulin expression system may provide a new approach to a controlled insulin gene therapy in skeletal muscle.

Animals ; Cell Line ; Dose-Response Relationship, Drug ; Doxycycline ; pharmacology ; Gene Expression Regulation ; drug effects ; Genetic Vectors ; genetics ; Insulin ; biosynthesis ; genetics ; Mice ; Muscle Fibers, Skeletal ; cytology ; metabolism ; Proinsulin ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Transfection

BACKGROUNDBmi-1 gene determines the proliferative capacity of normal and leukemia stem cells. Expression of Bmi-1 has been found in all types of myeloid leukemia cells in both humans and mice. This study aimed at assessing the effect of antisense Bmi-1 expression on K562 cells proliferation and p16 protein (p16) expression.

METHODSA transcriptional repressor, Bmi-1 cDNA was cloned by reverse transcriptase polymerase chain reaction (RT-PCR) of its mRNA from K562 cells. A plasmid expressing antisense Bmi-1 mRNA was then constructed by reverse design of PCR primers and cloned to the plasmid pLNCX2; G418 was added to the medium after the plasmid was successfully introduced in K562 cells by lipofectin-mediated DNA transfection. The effects of the antisense expression on the proliferation of K562 cells were analyzed by using microculture tetrazolium and colony forming. Cell cycle was analyzed by using flow cytometry. The p16 expression of K562 cells was observed by immunofluorescence histochemical stain.

RESULTSK562 cells transfected with antisense Bmi-1 plasmid grew significantly slower than that of controls (the parental K562 and cells transfected with empty plasmid). The colony forming ability of antisense Bmi-1 plasmid transfected cells decreased significantly (P < 0.01) compared with controls. The p16 expression of cells transfected with antisense Bmi-1 was upgraded more apparently than that of controls.

CONCLUSIONThe antisense Bmi-1 gene can inhibit the growth of K562 cell and upgrade expression of p16 in K562 cells.

Cell Cycle ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor p16 ; analysis ; Humans ; K562 Cells ; Nuclear Proteins ; antagonists & inhibitors ; genetics ; Plasmids ; Polycomb Repressive Complex 1 ; Proto-Oncogene Proteins ; antagonists & inhibitors ; genetics ; RNA, Antisense ; physiology ; Repressor Proteins ; antagonists & inhibitors ; genetics