Objective To explore the function of fluorescent probe JC-1 in detecting the changes of mitochondrial membrane potential(△?m)in early apoptotic cells.Methods After 2-ME was used to induce MUTZ-1 cell apoptosis,cells were dyed with fluorescent probe JC-1,and then the changes of △?m in the early stage of apoptotic cells were analyzed by flow cytometry or detected under fluorescent microscope. Results The control cells with high △?m are those forming JC-1 aggregates in the inner membrane of mitochondria,thus showing orange-red fluorescence.2-ME caused decrease of △?m in MUTZ-1 cells,in which JC-1 maintains monomeric form,thus showing only green fluorescence.The decreases of △?m were in a time-dependent manner,which were significantly higher than those in control group(P

Adult ; Dendritic Cell Sarcoma, Interdigitating ; complications ; surgery ; Humans ; Male ; Paraneoplastic Syndromes ; etiology ; Pemphigus ; etiology

Objective To explore a method for isolation and culture of human epidermal stem cells. Methods Epidermis was obtained by digesting human foreskin with Dispase Ⅱ and Trypsin-EDTA.After suspension on the epidermal stem cell medium (ESCM), these single epidermis cells were inoculated onto human collagen Ⅳ-coated flasks and cultured at 37 ℃ in a humidified atmosphere containing 5% CO_2 for 10 min. The nonadherent cells were rinsed off 10 min after inoculation, and the adherent cells continued to be cultured after enriching and abstraction by type Ⅳ collagen. The cell growth was observed through inverted microscope, and the cell cloning efficiency and time of clone sustain were also detected. Immunocytochemistry was used to observe the expression of ?_1-integrin and keratin 19(K19). Keratinocytes were served as controls. Results It was revealed by histological observation that colonies were formed 24 hours after inoculation. The isolated and cultured cell cloning efficiency was higher and the time of clone sustain was longer than that of the control group. Positive expression of ?_1-integrin and K19 of cultured cells was detected by immunocytochemistry. Conclusion Adult epidermal stem cells could be successfully isolated and cultured by adhension with type Ⅳ collagen and culture with ESCM.

This paper is in order to discussion with the composition and characteristics of Tibetan medicine plant resources, and promote the reasonable protection and utilization of the resources of Tibetan materia medica. Statistical analysis of species, distributions, and others of Chinese endemic seed plant from Tibetan medicine plants and usually used in the clinic of Tibetan medicine. The results showed that there are 523 species (25%) of Chinese endemic seed plant, belonging to 65 families and 162 genera, in about 2 000 varieties of Tibetan medicine plants recorded in relevant literatures. There are 180 Chinese endemic seed plant species (28%) belonging to 42 families and 72 genera from 625 medicine plants usually used in the clinic of Tibetan medicine. Specifically, the most of these Chinese endemic seed plant species are characteristic crude drug used in Tibetan medicine, and mainly or only distributed in Qinghai-Tibet Plateau. And a few species of them were intersected with traditional Chinese medicines (TCM) and other ethnic medicines. In addition, about 10% are listed in China Species Red List. The Qinghai-Tibet Plateau is the most abundant areas of Areal-types of the Chinese endemic seed plant. This is the biological and ecological reason formation the characteristics of Tibetan medicine plant resources. Therefore, strengthen the research of Chinese endemic seed plants used in Tibetan medicine is great significance for the reasonable protection and utilization of Tibetan medicine plant resources.
Drugs, Chinese Herbal ; chemistry ; Medicine, Tibetan Traditional ; Plants, Medicinal ; chemistry ; classification ; growth & development ; Seeds ; chemistry ; classification ; Tibet

The main difficulties and disputable problems on develping the national standard Standardized Manipulation of Acupuncture and Moribustion, Part 3: Auricular Acupuncture are debated in this paper from the three aspects of its meaning, scientific basis and key point. The difficult points mainly include manipulation standandization of auricular acupuncture, basis and science of the standards stipulated, selection of technical terms and definition, and why only the four kinds of manipulations are used for auricular acupuncture. Finally, the authors suggest that the Standardized Manipulation of Acupuncture and Moxibustion, Part 3: Auricular Acupuncture needs further to be studied and improved.
Acupuncture ; methods ; standards ; Acupuncture Points ; Acupuncture, Ear ; methods ; standards ; China ; Moxibustion ; methods ; standards

OBJECTIVETo study the effect of 2-methoxyestradiol(2-ME) on telomerase activity expression and apoptosis in human myelodysplastic syndrome cells line MUTZ-1.

METHODSMUTZ-1 cells were incubated with different concentrations of 2-ME, apoptosis rate and cell cycle were measured by flow cytometry (FCM). Telomerase activity in MUTZ-1 cells was examined by telomeric repeat amplification protocol-Enzyme linked immunosorbent assay (TRAP-ELISA).

RESULTSThe FCM analysis showed that cells in G0/G1 phase and S phase were decreased, while in G2/M phase increased after exposed to 1,2 and 4 micromol/L of 2-ME for 12 hours (P < 0.05). 1 and 2 micromol/L of 2-ME had no notable effect on MUTZ-1 cells as compared with the control group (P > 0. 05). Cells incubated with 1, 2 and 4 micromol/L of 2-ME for 36 hours were induced apoptosis, the percentage of apoptosis was between (12.87 +/- 0.86)% and (21.82 +/- 1.71)% with a dose- and time- dependent manner. Telomerase activity was significantly inhibited in these concentration and negatively correlated with cell number in G2/M phase (r = -0.979, P = 0.021) and increased apoptosis (r = -0.970, P = 0.030 ), respectively. Moreover, the inhibition effect of telomerase activity was enhanced in a dose- and time- dependent manner.

CONCLUSIONS2-ME-induced apoptosis and inhibition of telomerase activity provide a possible mechanism for explaining the 2-ME's anticancer activity.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line ; Estradiol ; analogs & derivatives ; pharmacology ; Humans ; Myelodysplastic Syndromes ; enzymology ; pathology ; Telomerase ; metabolism

To study the effects and possible mechanism of Vitamin K(2) (VK(2)) in the treatment of MDS-JSN04 cells, the changes of morphologic features of MDS-JSN04 cells were investigated by cytomorphology, the apoptosis of MDS-JSN04 cells was observed by transmission electron microscope; cellular proliferation was determined by the MTT assay; cell apoptosis, cell cycle shift and expression of myeloid-specific differentiation antigen (CD11b, CD13) were analyzed by flow cytometry (FCM). The expression of apoptosis-related genes bcl-2, survivin and bax were detected by retrotranscriptase polymerase chain reaction (RT-PCR); the activity of caspase-3 was determined by chemiluminescence assay. The results showed that the typical apoptotic morphological features appeared in cells treated with VK(2) for 72 hours; VK(2) induced apoptosis of MDS-JSN04 cells and in a dose-and-time-dependent manner, G(0)/G(1) cell arrest and significantly down-regulated the expression of bcl-2 and survivin, but had no effect on the expression of bax; the activity of caspase-3 significantly increased. It is concluded that VK(2) induces apoptosis of MDS-JSN04 cells through activating caspase-3 pathways and the apoptosis-related genes bcl-2, survivin may play an important role in this process.
Apoptosis ; drug effects ; CD11b Antigen ; analysis ; CD13 Antigens ; analysis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; Humans ; Inhibitor of Apoptosis Proteins ; Luminescent Measurements ; methods ; Microscopy, Electron, Transmission ; Microtubule-Associated Proteins ; genetics ; Myelodysplastic Syndromes ; genetics ; metabolism ; pathology ; Neoplasm Proteins ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Vitamin K 2 ; pharmacology ; bcl-2-Associated X Protein ; genetics

OBJECTIVETo evaluate transesophageal echocardiography (TEE)-guided periventricular device closure for treatment of ventricular septal defects (VSD) in children.

METHODSThe Amplatzer device was applied, the size of which was 1 or 2 mm larger than the VSD size assessed by TEE. Via a small lower sternotomy, the device was introduced through right ventricular surface to close the VSD under continuous TEE guidance without cardiopulmonary bypass.

RESULTThe procedures were successful in 6 cases, including 4 cases with muscular VSD and 2 cases with perimembranous VSD. The diameter of VSD was 3 mm to 10 mm and one case had pulmonary hypertension. One case of multi-muscular VSD with transposition of the great arteries failed to the treatment because the mitral open was interfered by the device. One case had bundle branch block in ICU stay and returned to normal without special therapy. The average operation time was 30 min and blood loss was 5 approximately 20 ml, no patients required blood transfusion; the average length of ICU stay was 24 h. Patients were discharged 1 to 2 d postoperatively. At a median follow-up for 12 months, all patients were asymptomatic and had no residual ventricular shunts.

CONCLUSIONWith the guide of TEE, intra-operative hybrid therapy is a safe and effective method for closure of muscular and perimembranous VSD without cardiopulmonary bypass in children.

Cardiac Catheterization ; methods ; Child ; Child, Preschool ; Echocardiography, Transesophageal ; Female ; Heart Septal Defects, Ventricular ; diagnostic imaging ; surgery ; Humans ; Infant ; Male ; Treatment Outcome

OBJECTIVETo in vitro study the inhibition effect and possible mechanism of As2S3 nanoparticles (As2S3 nano) on human MDS cell line MUTZ-1 and to compare with that of traditional As2S3.

METHODMUTZ-1 cells were treated with As2S3 nano and traditional regular-sized particles (TRSP) at different concentrations. The cell growth inhibition rate was determined by MTT assay, cell apoptosis by morphology and flow cytometry (FCM), cell cycle by FCM and the activity of caspase-3 by chemiluminescence assay.

RESULTSTreatment of As2S3 nano and TRSP at concentrations of 2, 4, 8 and 16 micromol/L for 48 h could lead to a significant dose-dependent decrease of MUTZ-1 cells and induce apoptosis. The percentages of inhibition were 48.9%, 75.9%, 89.4% and 96.5% in As2S3 nano vs 14.5%, 25.4%, 34.7% and 51.5% in TRSP and apoptosis rates were (12.9 +/- 1.9)%, (19.2 +/- 2.2)%, (30.1 +/- 2.5)% and (45.9 +/- 2.3)% in As2S3 nano vs (5.3 +/- 1.8%)%, (11.1 +/- 2.6)%, (19.3 +/- 2.3)% and (25.5 +/- 2.5)% in TRSP respectively. There was statistically significant difference in these two groups (P < 0.01). The proportion of cell in G2/M phase and the activity of caspase-3 of MUTZ-1 cells treated with A2S32 nano were significantly higher than those treated with control group and As2S3 TRSP groups (P < 0.01).

CONCLUSIONSAs2S3 nanoparticles and TRSP can inhibit the proliferation of MUTZ-1 cells and induce apoptosis, which maybe through activating caspase-3 pathways and increasing the proportion of G2/M phase. As2S3 nanoparticles can produce a much better antitumor effect than As2S3 TRSP do.

Apoptosis ; drug effects ; Arsenicals ; administration & dosage ; pharmacology ; Caspase 3 ; metabolism ; Cell Cycle ; drug effects ; Cell Line ; Cell Proliferation ; drug effects ; Humans ; Myelodysplastic Syndromes ; metabolism ; pathology ; Nanoparticles

OBJECTIVETo analyze occurrence, prevention and treatment of the complications of thoracoscopic assisted spine surgery.

METHODSRetrospective review of 182 patients who underwent standard thoracoscopic technique or video-assisted thoracic surgical procedure from October 1998 to August 2004. The treatment of thoracic diseases included debridement, decompression with (or) reconstruction. The total number of complications were recorded, and its mechanism, prevention and treatment were analyzed.

RESULTSComplications occurred in 16 patients, 12 cases of perioperative complications included 3 patients suffered from pneumonia, 3 pulmonary atelectasis, 2 patients' lung injured by trocar, 1 patient obtained transient monoplegia, 2 suffered from transient intercostal nerve pain and 1 had superficial incision infection. Long-term complications occurred in 4 cases: spinal tuberculosis relapsed 2 cases (one who had diabetes obtained relapse in 8 months of post-operation and another relapsed with complex spinal tuberculosis in 4 weeks postoperation), 2 patients suffered from kyphosis deformity and pain.

CONCLUSIONSThe type and incidence of complications with thoracoscopic spine surgery mainly depend on indication, operation procedures and anesthesia, only by limit surgical indication, ameliorate technique, obey surgical principle and consummate perioperative treatment can we obtain mini-invasive effect by thoracoscopic assisted spine surgery.

Adolescent ; Adult ; Decompression, Surgical ; adverse effects ; methods ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Spinal Diseases ; surgery ; Spinal Fusion ; adverse effects ; methods ; Thoracic Vertebrae ; surgery ; Thoracoscopy ; adverse effects