OBJECTIVETo compare the efficacy on vertebrobasilar insufficiency (VBI) between auricular acupuncture therapy and oral administration of medicine.

METHODSSixty patients of VBI were randomized into an auricular acupuncture therapy group and a medicine group, 30 cases in each one. In the auricular acupuncture group, acupuncture was applied bilaterally to gan (CO12) and jiejie (HX8) on the ears and needles were retained for 15 min. After needle withdrawal, the vaccariae semen were fixed with plaster at naogan (AT3, 4i), zhen (AT3), jing (AH12), shen (CO10) and pi (CO13) on the ears. In the medicine group, flunarizine hydrochloride capsules (Sibelium), 5mg were prescribed for oral administration, once every night. The treatment lasted continuously for 2 weeks (14 days) in the two groups. In 2 weeks, the clinical efficacy was assessed and the transcranial doppler (TCD) examination was performed.

RESULTSAfter treatment, the symptom scores were all apparently reduced in the patients of the two groups (P < 0.01, P < 0.05). Compared with the medicine group, the reduced score was much more obvious in the auricular acupuncture group (P < 0.05), indicating the significant difference. After treatment, with TCD examination, the blood velocity was increased to different degrees in the patients of low velocity type in the auricular acupuncture group and the medicine group; that was reduced to different degrees in the patients of high velocity type in the auricular acupuncture group and the medicine group. All of them were different significantly as compared with those before treatment (all P < 0.05). But the difference was not significant between the two groups (both P > 0.05). In comparison of clinical efficacy between the two groups, the effective rate was 93.3% (28/30) in the acupuncture group and better than 76.7% (23/30) in the medicine group, indicating the significant difference in comparison (P < 0.05).

CONCLUSIONThe auricular acupuncture therapy achieves the definite efficacy on VBI and the efficacy is better than flunarizine hydrochloride capsules.

Acupuncture Points ; Acupuncture, Ear ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Vertebrobasilar Insufficiency ; therapy

OBJECTIVETo study the pollution status of Legionella species in hot spring vacation center and the related factors.

METHODSField surveys were performed in four big hot spring vacation centers of Changping district. Uniform questionnaires was used and colony count was made together with the isolation of Legionella species from hot spring water based on mip gene typing.

RESULTS47 isolates of Legionella pneumophila (Lp) from 87 samples showed 4 serotypes as Lp1, Lp6, Lp12, Lp5 with percent of 57.45%, 21.28%, 14.89%, 6.38% respectively. The hot spring centers controlled the temperature of recycled water between 34-47 degrees C by hot water heating and filtrating system. All the isolates were cultured from the hot water with temperature between 34-44 degrees C: 56.75% (21/37) in high temperature (40-47 degrees C) and 61.90% (26/42) in low temperature (34-39.9 degrees C). There were no statistically significant difference between the high and the low temperature samples (P > 0.05). In the four hot spring vacation centers, the pH value was under control at 6.4-7.3 and the ambient temperature was under control between 26-28 degrees C. The humidity was controlled between 56% -69% relative humidity, which were the best growing conditions for the Legionella species. Disinfectors as chlorine deviratives was used in the four hot spring vacation centers. Though the concentration of chlorine in the water was 0.3-0.5 mg/L, 14.29%-48.00% of the samples were still positive of having Legionella species.

CONCLUSIONThe pollution of Legionella species was considered to be quite serious in the four hot spring vacation centers and the predominant serotype was Lp1. The pH and temperature of the hot spring water, ambient temperature and humidity and the way of heating up the water were the best conditions for the growth of Legionella species in these centers. Because of the high temperature of the hot spring water, chlorine of the disinfector volatilized quickly, affecting the effect of disinfection. The result revealed that water temperature achieving 44 degrees C could have had the effect of prevention.

China ; Disinfection ; Environmental Monitoring ; Hot Springs ; microbiology ; Legionella ; growth & development ; isolation & purification ; Temperature ; Travel ; Water Microbiology

Actins ; metabolism ; Carcinoembryonic Antigen ; metabolism ; Desmin ; metabolism ; Diagnosis, Differential ; Epithelial Cells ; metabolism ; pathology ; Follow-Up Studies ; Humans ; Kidney Neoplasms ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Neoplasms, Complex and Mixed ; metabolism ; pathology ; surgery ; Neoplasms, Glandular and Epithelial ; metabolism ; pathology ; surgery ; Stromal Cells ; metabolism ; pathology ; Vimentin ; metabolism

OBJECTIVETo investigate apoptosis of tumor infiltrating dendritic cells (TIDC) and their expression of Fas/FasL (CD95/CD95L) in human endometrioid adenocarcinoma.

METHODSThe apoptotic rate of TIDC was measured in 45 cases of endometrioid adenocarcinoma and 20 cases of normal endometrium tissues (control) by double-label immunohistochemistry using the monoclonal antibody S-100 protein and TUNEL technique. The expressions of Fas and FasL in TIDCs were detected using double-label immunohistochemistry and imaging analysis.

RESULTSThe apoptotic rate of TIDCs in endometrioid adenocarcinoma were significantly higher than that in normal endormetrium [(13.02∓0.64)% vs (6.82∓0.53)%, P<0.05]. The expression levels of Fas in the TIDCs were significantly lower, whereas FasL expression significantly higher in endometrioid adenocarcinoma than in normal endormetrium (7.88∓1.05 vs 19.25∓3.03, P<0.05; 12.95∓2.25 vs 7.51∓1.14, P<0.05).

CONCLUSIONIncreased apoptosis of the TIDCs and abnormal expression of Fas/FasL in TIDCs in endometrioid adenocarcinoma may lead to tumor immune escape.

Apoptosis ; physiology ; Carcinoma, Endometrioid ; immunology ; metabolism ; pathology ; Case-Control Studies ; Dendritic Cells ; immunology ; Endometrial Neoplasms ; immunology ; metabolism ; pathology ; Fas Ligand Protein ; genetics ; metabolism ; Female ; Humans ; Lymphocytes, Tumor-Infiltrating ; immunology ; Tumor Escape ; fas Receptor ; genetics ; metabolism

This study was purposed to investigate the inhibition of hTERT antisense oligodeoxynucleotide (ASODN) on the proliferation and telomerase activity in HL-60 cells and to explore the relativity between the telomerase activity and the expression of hTERT gene in HL-60 cells. After treated by hTERT ASODN the expression of hTERT was detected by RT-PCR, the morphological changes of HL-60 cells was observed with inverted microscopy, the cell proliferation was measured by MTT method, and the telomerase activity was determined with TRAP-ELISA and TRAP-PAGE. The results showed that after sealing hTERT gene with ASODN for 72 hours, the expression of hTERT gene was significantly inhibited, the cell growth was repressed and the ability of proliferation decreased, and the effect was specific in sequence and dependent in dose and time. OD(450-690) values were 2.648 +/- 0.42, 1.504 +/- 0.47, 1.223 +/- 0.39, 0.944 +/- 0.16 respectively, as the cells were treated with 0, 10, 20, 30 micromol/L ASODN for 72 hours. The difference was significant as compared 10, 20, 30 micromol/L groups with 0 micromol/L ASODN group respectively (P < 0.05), but the difference was no significant when compared 20 micromol/L SODN group (2.376 +/- 0.65) with untreated group (2.648 +/- 0.42) (P > 0.05). TRAP-PAGE detection revealed that comparing ASODN groups with SODN groups the telomerase image bands were decreased and least was found in groups of 30 +/- mol/L. It is concluded that the hTERT ASODN may inhibit the proliferation and down-regulate the telomerase activity in HL-60 cells by sealing the expression of hTERT gene.
Cell Proliferation ; drug effects ; HL-60 Cells ; Humans ; Oligonucleotides, Antisense ; biosynthesis ; genetics ; Telomerase ; biosynthesis ; genetics ; metabolism ; pharmacology ; Transfection

OBJECTIVETo characterize the time course of spontaneous differentiation of in vitro cultured human embryonic stem cells (hESCs) into hematopoietic cells to provide experimental evidence for induction of hematopoietic commitment of hESCs.

METHODSIn human embryoid bodies (hEBs) derived from spontaneous differentiation of chESC3, a hESC cell line we established previously, the expressions of such genes as KDR, Bmi1, Scl and gata2 were detected by RT-PCR every other day during the 12-day differentiation to monitor the process of the hematopoiesis. The hematopoietic stem cell marker CD34 was examined using flow cytometry to evaluate the efficiency of hematopoietic differentiation of the cells on days 6, 8, 10 and 12. The spontaneously differentiated hESCs were seeded in the hematopoietic colony culture system to study the hematopoietic colony forming ability. Immunocytochemical staining for CD45 was performed on the hEBs to examine the emergence of mature hematopoietic cells.

RESULTSThe expressions of the hematopoietic stem cell-related genes KDR and Bmi-1 were detected in the hESCs, and on days 4 to 6, the two genes were upregulated with prolonged cuture of the hEBs. Scl and gata2 gene expressions were detected since 6-8 days of culture and maintained high expressions till day 12. Flow cytometry revealed a gradual increase in CD34-positive cells in the culture, with positivity rates on days 6, 8, 10, and 12 of (1.4-/+0.4)%, (3.4-/+1.3)%, (5.5-/+2.2)%, and (5.1-/+1.7)%, respectively. The numbers of CD43-positive cell colonies on days 6, 8, 10, and 12 were 0, 7-/+2, 37-/+11, and 89-/+29 in each 10(5) cells, respectively. Immunocytochemical staining identified CD45-positive cells on days 10, 12, 15, and 18 in the cell colonies, with the positive cell numbers of 0, 40.5-/+15.09, 178.6-/+55.89, and 253.0-/+52.04, respectively.

CONCLUSIONThe hESCs undergo spontaneous hematopoietic differentiation in 3 stages, including the differentiation into germ layer-specific cells (days 6-8), expansion period of the hematopoitic progenitors (days 8-12), and maturation of the hematopoietic cells (after day 15).

Animals ; Antigens, CD34 ; metabolism ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; Cell Culture Techniques ; Cell Differentiation ; Embryonic Stem Cells ; cytology ; metabolism ; GATA2 Transcription Factor ; genetics ; Gene Expression Regulation ; Hematopoietic Stem Cells ; cytology ; metabolism ; Humans ; Mice ; Nuclear Proteins ; genetics ; Polycomb Repressive Complex 1 ; Proto-Oncogene Proteins ; genetics ; Repressor Proteins ; genetics ; T-Cell Acute Lymphocytic Leukemia Protein 1 ; Time Factors

OBJECTIVETo investigate the effect of vascular bundle implantation in autogenous bone graft on angiogenesis.

METHODSThirty-six New Zealand white rabbits were evaluated in this study. A portion of bilateral radial bones of a rabbit were removed as free bone grafts, whose periostea were peeled off. In test group, the external maxillary artery bundle was passed through the marrow cavity of the bone. In control group, there was no vascular bundle implantation. Each bone was placed in masseter muscle separately. The rabbits were sacrificed and the specimens were procured at 3 days, 1, 2, 3, 4 and 6 weeks after surgery for histological observation, Chinese ink perfusion and CD34 immunohistochemistry. Microvessel density (MVD) was assessed in order to evaluate angiogenesis of autogenous bone grafts.

RESULTSThe bone grafts were found revascularization in 3 days after surgery in the test group, whereas at 2 weeks in the control group. In 3 days, 1 week, 2 weeks, 3 weeks and 4 weeks after surgery, the MVD of test group was significantly higher than that of control group. In 4 weeks after surgery, angiogenesis of test group reached to peak.

CONCLUSIONVascular bundle implantation improved angiogenesis in non-vascularized autogenous bone graft in this study.

Animals ; Bone Transplantation ; Bone and Bones ; Prostheses and Implants ; Rabbits

OBJECTIVEIt is well documented that epilepsy can increase neurogenesis in certain brain regions and cause behavioral alternations in patients and different epileptic animal models. A series of experimental studies have demonstrated that neurogenesis is regulated by various factors including glucocorticoid (CORT), which can reduce neurogenesis. Most of studies in animal have been focused on adulthood stage, while the effect of recurrent seizures to immature brain in neonatal period has not been well established. This study was designed to investigate how the recurrent seizures occurred in the neonatal period affected the immature brain and how CORT regulated neurogenesis in immature animals.

METHODSNeonatal rats were subjected to 3 pilocarpine-induced seizures from postnatal day 1 to day 7. Then neurogenesis at different postnatal ages (i.e. P8, P12, P22, P50) was observed. Behavioral performance was tested when the rats were mature (P40), and plasma CORT levels following recurrent seizures were simultaneously monitored.

RESULTSRats with neonatal seizures had a significant reduction in the number of Bromodeoxyuridine (BrdU) labeled cells in the dentate gyrus compared with the control groups when the animals were euthanized on P8 or P12 (P<0.05); whereas there was no difference between the two groups on P22. Until P50, rats with neonatal seizures had increased number of BrdU-labeled cells compared with the control group (P<0.05). In Morris water maze task, pilocarpine-treated rats were significantly slower than the control rats at the first and second day, and there were no differences at other days. In probe trial, there was no significant difference in time spent in the goal quadrant between the two groups. Endocrine studies showed a correlation between the number of BrdU positive cells and the CORT level. Sustained increase in circulating CORT levels was observed following neonatal seizures on P8 and P12.

CONCLUSIONNeonatal recurrent seizures can biphasely modulate neurogenesis over different time windows with a down-regulation at early time and up-regulation afterwards, cause persistent deficits in cognitive functions of adults, and increase the circulating CORT levels. CORT levels are related with the morphological and behavioral consequences of recurrent seizures.

Age Factors ; Animals ; Animals, Newborn ; Critical Period (Psychology) ; Dentate Gyrus ; cytology ; growth & development ; metabolism ; Glucocorticoids ; blood ; Male ; Maze Learning ; physiology ; Neurons ; cytology ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Seizures ; metabolism ; pathology ; physiopathology ; Stem Cells ; cytology ; metabolism

OBJECTIVETo establish an accurate and efficient method for detecting recent thymic output function and analyze the content of T-cell receptor (TCR) rearrangement excision circles (TRECs) within peripheral blood mononuclear cells (PBMCs).

METHODSAccording to the specific sequence of TCRdelta, the primers and the fluorescent probe (TaqMan) were designed and synthesized. The standard quantitative template was constructed by T/A cloning. The method for detecting TRECs was established after optimization of reaction condition, then its specificity, sensitivity and stability were tested. Quantitative detection of TRECs in DNA of PBMCs from normal individuals and patients of chronic hepatitis B were preformed by real-time PCR using TaqMan technique.

RESULTSDetection of TRECs was quick and accurate by real-time fluorescence quantitative PCR. The CV value of Ct was 1.06%, the product was specific which was confirmed by electrophoresis and sequencing and the method showed high sensitivity. The mean value of TRECs from normal individuals was (7767.4 +/- 2369.5) copies/10(6)PBMCs in healthy controls at age 21.45 but (28,374.4 +/- 7820.4) copies/10(6)PBMCs in those at age 16.20 (P < 0.05). The mean value of TRECs from patients with chronic hepatitis B was (6480.9 +/- 2031.2) copies/10(6) PBMCs in those at age 21.45, which was statistically significant as compared with normal individuals at age 21.45.

CONCLUSIONReal-time fluorescence quantitative PCR for detecting the TRECs is an accurate, efficient and stable method and the recent thymic output function might decrease in patients with chronic hepatitis B.

Adult ; DNA Primers ; Female ; Gene Rearrangement, T-Lymphocyte ; Hepatitis B, Chronic ; blood ; genetics ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Receptors, Antigen, T-Cell, gamma-delta ; genetics ; Reproducibility of Results ; Thymus Gland ; immunology ; metabolism ; Young Adult

OBJECTIVETo investigate the effect of hTERT antisense oligodeoxynucleotide (ASODN) on the oncogenicity and the inductive apoptosis of HL-60 cells.

METHODSApoptosis of HL-60 cells was detected by flow cytometry (FCM) and agarose gel electrophoresis. Both treated and untreated HL-60 cells were collected and transplanted into 5 BALB/c nude mice respectively, the formation of transplanted neoplasm and its morphologic change were observed. After the transplanted neoplasms were uniform with the ameliorated method in another 10 BALB/c nude mice, they were divided into 2 groups and injected ASODN and PBS into the neoplasm respectively. Seven days later, the tumor were measured, its morphology were observed, and the apoptotic cells were detected with a TUNEL kit.

RESULTSAfter 72 h treatment there were DNA ladders and early apoptosis peak in hTERT ASODN treated HL-60 cells but was none in SODN treated and blank control cells. In tumor formation experiment, neoplasms were formed in ASODN treated group at 16-17 d and untreated group at 12-13 d. Neoplasm was formed in 2 of 5 ASODN treated mice and 4 of 5 untreated mice respectively. In untreated mice tumor tissues were rich in blood vasa and stromal tissue compared with that in ASODN treated mice. In tumor therapy experiment, before treatment, there was no difference in the average neoplasm physical volume between ASODN treated group [(100.9 +/- 24.6) mm3] and PBS treated group [(98.4 +/- 23.1) mm3] (P > 0.05). After treatment, the neoplasm volume in ASODN treated group [(422.7 +/- 326.4) mm3] was smaller than that in PBS treated group [(786.4 +/- 357.6) mm3] (P < 0.05). Histologically, there were many apoptosis cells in ASODN treated group, but was seldom seen in PBS treated group. The TUNEL positive cells in ASODN treated group were much more than that in PBS treated group (P < 0.05).

CONCLUSIONThe hTERT ASODN induces apoptosis of HL-60 cells in vitro, reduces the tumor formation in BALB/c nude mice and inhibits the growth of the transplanted neoplasm.

Animals ; Apoptosis ; drug effects ; HL-60 Cells ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Oligodeoxyribonucleotides, Antisense ; pharmacology ; Telomerase ; genetics ; Transfection ; Xenograft Model Antitumor Assays