Diagnostic Errors ; Female ; Humans ; Middle Aged ; Thyroid Nodule ; diagnostic imaging ; Ultrasonography ; Zenker Diverticulum ; diagnostic imaging

Objective To investigate the mechanism and suppression effect of human cytomegalovirus (HCMV) on proliferation of megakaryocyte progenitors(CFU-Mk)in vitro.Methods Colony forming unit-assay was applied to observe the effect of HCMV-AD 169 strain on CFU-Mk of cord blood. The technique of PCR was used to demonstrate the existence of HCMV-AD 169 DNA in the colony cells of cultured CFU-Mk.Results 1.The number of CFU-Mk colonies in HCMV-infected groups decreased significantly compared with that of control group. The CFU-Mk formation was inhibited significantly after HCMV-AD 169 strain infection.The suppression effect showed a dose-dependent fashion: 46.7 % inhibition with 10 -1of HCMV, 29.7 % with 10 -2 and 14.5 % with 10 -3 in the CFU-Mix assay. The peak of CFU-Mk colonies (d16-18) was not significantly different between control group and experimental groups, but the duration of the CFU-Mk colonies in infected groups was significantly shorter than that in control group.2. HCMV-DNA was positively detected in the colony cells of viral infected group by PCR, while negative in control group.Conclusions HCMV-AD 169 strain may inhibit the differentiation and proliferation of CFU-Mk by infecting the hematopoietic progenitors. HCMV may cause the suppression of hematopoiesis by direct infection, which may be the main reason for HCMV infection associated with thrombocytopenia.

OBJECTIVETo construct the human/mouse chimeric antibody of a functional anti-death receptor 5 (DR5) antibody. Methods The viable region of light chain (VL) and viable region of heavy chain (VH) genes of anti-DR5 antibody were amplified and cloned into the light- and heavy-chain expression vectors respectively, then the recombinant plasmids were co-transfected into dihydrofolate reductase(-) Chinese hamster ovary cell (CHO-dhfr(-)) for expression. The positive clone was screened by the two selective genes (neo and dhfr). The humanization and specificity of chimeric antibody was identified by ELISA and Western blotting, and the tumoricidal activity of the expressed chimeric antibody was detected by tetrazolium salt phenazine methosulfate assay.

RESULTSThe expression vectors stably expressed chimeric antibody in CHO-dhfr(-). In the cell supernatant of the F4' clone, the human IgG heavy constant region and light constant region were identified. Moreover, the secreted chimeric antibody retained the binding capacity to the antigen (DR5) and decreased the cell viability of Jurkat and HCT116 cells to 73.15% and 77.30% in vitro respectively.

CONCLUSIONThe human/mouse anti-DR5 chimeric antibody has been successfully expressed in eukaryotic cells and shows tumoricidal activity, which establishes a foundation for the future research of humanized antibody medicine.

Animals ; Antibodies ; genetics ; immunology ; pharmacology ; Antineoplastic Agents ; immunology ; pharmacology ; CHO Cells ; Cell Survival ; drug effects ; Cricetinae ; Cricetulus ; Gene Expression ; Humans ; Mice ; Protein Engineering ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; genetics ; immunology ; Recombinant Fusion Proteins ; genetics ; immunology ; pharmacology

A large amount of nicotinamide adenine dinucleotide phosphate (NADPH) is required for fatty acid synthesis and maintenance of the redox state in cancer cells. Malic enzyme 1(ME1)-dependent NADPH production is one of the three pathways that contribute to the formation of the cytosolic NADPH pool. ME1 is generally considered to be overexpressed in cancer cells to meet the high demand for increased de novo fatty acid synthesis. In the present study, we found that glucose induced higher ME1 activity and that repressing ME1 had a profound impact on glucose metabolism of nasopharyngeal carcinoma(NPC) cells. High incorporation of glucose and an enhancement of the pentose phosphate pathway were observed in ME1-repressed cells. However, there were no obvious changes in the other two pathways for glucose metabolism: glycolysis and oxidative phosphorylation. Interestingly, NADPH was decreased under low-glucose condition in ME1-repressed cells relative to wild-type cells, whereas no significant difference was observed under high-glucose condition. ME1-repressed cells had significantly decreased tolerance to low-glucose condition. Moreover, NADPH produced by ME1 was not only important for fatty acid synthesis but also essential for maintenance of the intracellular redox state and the protection of cells from oxidative stress. Furthermore, diminished migration and invasion were observed in ME1-repressed cells due to a reduced level of Snail protein. Collectively, these results suggest an essential role for ME1 in the production of cytosolic NADPH and maintenance of migratory and invasive abilities of NPC cells.
Carcinoma ; Cell Line, Tumor ; Cell Movement ; Cell Survival ; Glucose ; metabolism ; Glycolysis ; Humans ; Malate Dehydrogenase ; metabolism ; NADP ; metabolism ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Invasiveness ; Oxidation-Reduction ; Oxidative Phosphorylation ; Pentose Phosphate Pathway ; Proto-Oncogene Proteins c-akt ; metabolism

OBJECTIVETo assess the role of probiotics in the prevention of neonatal necrotizing enterocolitis (NEC) and to investigate the risk factors for NEC.

METHODSA total of 2528 hospitalized neonates between January 2002 and May 2005 were assigned into either receiving prophylactic use of probiotics bifoco (Prevention group, n=1182) or without probiotics supplementation (Control group, n = 1346). The incidence of NEC was compared between the two groups. The risk factors for NEC were investigated by conditional logistic regression multifactorial analysis.

RESULTSThere were 19 cases of NEC in the Control group (1.41%), but only 6 cases in the Prevention group (0.51%) (P < 0.05). Gestational age (OR = 5.521), hypoxicdouble ended arrowischemic encephalopathy (OR = 3.887), specticemia (OR = 4.854) and critical illness scores (OR = 5.989) were the risk factors for NEC, while the prophylactic use of probiotics was an independent protective factor for NEC (OR = 0.255).

CONCLUSIONSThe prophylactic use of probiotics may reduce the incidence of NEC in neonates.

Case-Control Studies ; Enterocolitis, Necrotizing ; prevention & control ; Female ; Humans ; Infant, Newborn ; Male ; Probiotics ; therapeutic use

AIMTo determine the effect of piperazinyl estrone, a new estrogen derivative, on bone turnover, bone mass and uteri in ovariectomized rats.

METHODSFemale Sprague-Dawley rats were ovariectomized (OVX) or sham operated (sham) at the age of 3 months and treated with estrone (E) at 0.75 mg.kg-1.d-1, or with piperazinyl estrone (P-E) at 1 or 10 mg.kg-1.d-1, orally, for 3 months. At the time of death, the uterine weight was measured. Bone histomorphometric analysis of proximal tibial metaphyses (PTM) was performed in undecalcified sections.

RESULTSBone histomorphometric data showed that the percent trabecular area (% Tb.Ar) of OVX rats with bone high turnover was significantly decreased. The uteri were atrophied. The percent trabecular area (% Tb.Ar) of estrone treated group was increased in decreasing bone turnover manner. But the size and weight of uteri in this group were increased vs OVX group. The bone loss induced by OVX was preserved by P-E treatment, but the mechanism of maintaining bone is different from that of E-treated rats. P-E treatment in low dose did not decrease any bone formation indices, such as percent labeling perimeter, bone formation rate per bone volume (BFR/BV), except bone mineral apposition rate (MAR) compared with E-treated group, and maintained them at OVX level. The uteri were found to be in atrophy compared with the match dose (0.75 mg) of E-treated OVX rats. But rats treated with high dose of P-E showed the same change like E-treated group.

CONCLUSIONThe finding of this study shows that lower dosage of piperazinyl estrone has effect on preventing the bone losses in OVX rats, while the bone formation and the uterus are not affected, thus supporting the hypothesis that piperazinyl estrone has the potential to prevent postmenopausal bone loss in women with less side effects.

Animals ; Atrophy ; prevention & control ; Bone Density ; Estradiol Congeners ; pharmacology ; therapeutic use ; Estrone ; analogs & derivatives ; pharmacology ; therapeutic use ; Female ; Organ Size ; drug effects ; Osteogenesis ; drug effects ; Osteoporosis ; prevention & control ; Ovariectomy ; Rats ; Rats, Sprague-Dawley ; Uterus ; pathology

Objective To explore the intervention effect and mechanism of sitagliptin on renal injury in type 1 diabetic (T1DM) mice.Methods The modeling group was intraperitoneally injected with streptozotocin (150 mg· kg-1) once and blood glucose ≥ 16.7 mmol · L-1 was considered as diabetes mellitus (DM).DM mice were randomly divided into two groups:model group and experimental group.Experimental group was treated with sitagliptin (15 mg · kg-1 · d-1) via intragastric administration for 4 weeks while the mice in model group and normal group treated with equal volume of ultrapure water.The 24 h microalbuminuria(ALB) was determined after administration.At the end of the experiment,urea nitrogen (BUN) and creatinine (SCr) in serum was detected.The protein expression levels of transforming growth factor-β1 (TGF-β1),Smad2,Smad3 were measured by Western blot.Results After administration sitagliptin for 4 weeks,the contents of ALB in model group and experimental group were (11.96 ± 3.36) (2.46 ±0.97) mg/24 h with significantly(P ＜0.001).The index of kidney weight/body weight(KW/BW) in normal group,model group and experimental group were 15.20 ±2.24,21.43 ±2.16,15.14 ±4.14;compared with normal group,the difference was significantly increased (P ＜ 0.05);compared with model group,the difference was significantly increased (P ＜0.05).The SCr in model group and experimental group were (352.58 ±47.09),(238.51 ± 53.03) μmol · L-1;the BUN in the two groups were(26.08 ±4.65),(10.40 ±2.47)mmol · L-1;compared with model group,the difference was significantly increased (P ＜0.01,P ＜0.001).The expression levels of TGF-β1,Smad2/3 and p-Smad2/3 in normal group,model group and experimental group were 0.19 ±0.02,0.12 ±0.02,0.07 ±0.01;0.23 ±0.02,0.27 ±0.04,0.13 ±0.01;0.18 ±0.01,0.14 ±0.01,0.11 ±0.00,compared with normal group,the difference was significantly increased (P＜0.05,P＜0.01);compared with model group,the difference was significantly increased (P ＜0.05,P ＜0.01).Conclusion Experimental delaying the progression of T1DM nephropathy without not decreasing blood glucose can effect partly through inhibiting TGF-β1/Smad2/3 pathway.

OBJECTIVETo develop quality of life questionnaire of Chinese medicine for postoperative patients with colorectal cancer (QLQ-CMPPCC), thus comprehensively and objectively evaluating the clinical efficacy of Chinese medicine and pharmacy in treating postoperative patients with colorectal cancer (CC).

METHODSThe theoretical structure model of the questionnaire was addressed in combined with basic theories of Chinese medicine according to the principle of WHO quality of life (QOL). The primary questionnaire was developed using methods of structuralization policy making after we extensively retrieve various universal and specific questionnaires for CC cancer patients at home and abroad. The 205 CC patients were tested by questionnaire. The items were screened using experts grading method, item selection analysis, dispersion trends of standard deviation, t-test, correlation coefficient method, factor analysis,and Cronbach's alpha.

RESULTSThe QLQ-CMPPCC was developed containing four domains of physical, psychological, independence, and social functions, involving 20 aspects and 54 items. Of them, non-fistula patients answered 43 items and fistula patients answered 46 items. One item covered the general QOL evaluation.

CONCLUSIONSQLQ-CMPPCC showed Chinese medical features. It comprehensively reflected the connotation of QOL for postoperative CC patients. It could be taken as a tool for evaluating Chinese medical efficacy for postoperative CC patients.

Colorectal Neoplasms ; surgery ; Humans ; Medicine, Chinese Traditional ; methods ; Postoperative Period ; Quality of Life ; Surveys and Questionnaires ; Treatment Outcome

AIMTo record funny currents (If) of ventricular myocytes and to analysize hyperpolarization-activated cation channel(HCN) expression in the rats of different ages.

METHODSFresh ventricular myocytes were isolated from 3 days rats and adult rats.HCN expressions were measured by real-time quantitative polymerase chain reaction(real-time PCR). It was recorded through whole-cell patch clamp.

RESULTSHCN1, HCN2, HCN3, HCN4 mRNA represented 0.23% +/- 0.01%, 83.58% +/- 0.04%, 0.79% +/- 0.01%, 15.44% +/- 0.01% of total HCN mRNA in the neonatal rats, respectively. If was recorded and the threshold for activation was -75 mV. In the adult rat, HCN1, HCN2, HCN3, HCN4 mRNA represented 0.72% +/- 0.02%, 91.58% +/- 0.08%, 0.27% +/- 0.02%, 7.12% +/- 0.02% of total HCN mRNA. The ratio of HCN2 to HCN4 was approximately (13.06 +/- 0.21):1. The threshold for activation of If was approximately -115 mV in the adult rats.

CONCLUSIONWith the development of rats, the value of If is smaller. The threshold for activation of If is more negative. The ratio of HCN2 to HCN4 is bigger.

Age Factors ; Animals ; Animals, Newborn ; Cells, Cultured ; Cyclic Nucleotide-Gated Cation Channels ; metabolism ; physiology ; Heart Ventricles ; cytology ; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels ; Ion Channels ; metabolism ; Myocytes, Cardiac ; cytology ; physiology ; Patch-Clamp Techniques ; Potassium Channels ; metabolism ; physiology ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo analysis the effect of amiodarone on funny current (I(f)) and hyperpolarization-activated cation channel (HCN) gene expressions of the neonatal rat ventricular myocytes.

METHODSVentricular myocytes of 1 - 3 days-old rats were isolated and cultured. The cardiomyocytes were treated by amiodarone (0.01, 0.1, 1, 10, 100 micromol/L) for 3 hours or amiodaron (10 micromol/L) for 0, 0.5, 1, 3, 6 hours. The I(f) and HCN 1 - 4 gene expressions were measured through the whole-cell configuration of the patch-clamp technique and real-time quantitative polymerase chain reaction (real-time PCR) using SYBR Green PCR kit.

RESULTS(1) HCN1, HCN2, HCN3 and HCN4 represented (0.23 +/- 0.01)%, (83.58 +/- 0.04)%, (0.79 +/- 0.01)% and (15.44 +/- 0.01)% of total HCN mRNA, respectively. (2) Amiodaron resulted in a dose-dependent I(f) [(3.1 +/- 0.9)%, (9.7 +/- 2.4)%, (36.7 +/- 5.8)%, (80.3 +/- 1.8)% and (85.9 +/- 3.1)%, respectively at -145 mV, IC(50) (1.32 +/- 0.28) micromol/L], HCN2 [(2.1 +/- 0.8)%, (8.9 +/- 3.6)%, (30.1 +/- 4.2)%, (78.3 +/- 3.6)% and (81.1 +/- 1.9)%, respectively] and HCN4 decrease [(0.5 +/- 0.2)%, (2.1 +/- 2.6)%, (8.8 +/- 3.2)%, (60.1 +/- 4.6)% and (59.6 +/- 6.5)%, respectively]. (3) Amiodaron (10 micromol/L) also induced a time-dependent I(f) [(1.1 +/- 0.1)%, (12.6 +/- 2.3)%, (80.6 +/- 2.2)% and (80.1 +/- 2.1)%, respectively], HCN2 [(1.0 +/- 0.1)%, (9.8 +/- 3.9)%, (82.9 +/- 4.6)% and (83.9 +/- 1.7)%, respectively] and HCN4 decrease [(0.1 +/- 0.1)%, (1.9 +/- 1.1)%, (59.4 +/- 7.8)% and (60.9 +/- 3.1)%, respectively]. However, HCN1 and HCN3 expressions were not affected by amiodaron treatment.

CONCLUSIONCurrent density of I(f) and the expression of HCN2 and HCN4 were decreased by amiodaron which might be the possible antiarrhythmic working mechanisms of amiodaron.

Amiodarone ; pharmacology ; Animals ; Animals, Newborn ; Cells, Cultured ; Cyclic Nucleotide-Gated Cation Channels ; genetics ; metabolism ; Female ; Gene Expression ; Heart Ventricles ; metabolism ; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels ; Male ; Myocytes, Cardiac ; drug effects ; metabolism ; Patch-Clamp Techniques ; Potassium Channels ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley