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.

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

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

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 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

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

The study was aimed to investigate the mechanism of proliferation inhibition and apoptosis of MDS-RAEB MUTZ-1 cells induced by 2-methoxyestradiol (2-ME), the cell proliferation was determined by MTT assay, apoptosis rate was determined with annexinV-FITC/PI double staining and cell cycle was analyzed by flow cytometry (FCM) after MUTZ-1 cells were treated with different concentrations of 2-ME; the changes of morphologic features of MUTZ-1 cells were observed with Wright-Giemsa's staining; lactate dehydrogenase was determined by Beckman Counter; and agarose gel electrophoresis was used to verify whether 2-ME can induce apoptosis of MUTZ-1 cells. The results showed that 2-ME inhibited the proliferation of MUTZ-1 cells in a dose-and time-dependent manner and caused a sustained arrest at G(2)/M phase in MUTZ-1 cells; the typical apoptotic morphological features appeared in MUTZ-1 cells; the production of lactate dehydrogenase was up-regulated and the marked DNA ladder pattern of internucleosomal fragmentation was observed. It is concluded that the mechanism of proliferation inhibition and apoptosis of MUTZ-1 cells induced by 2-ME is probably related with the G(2)/M cell cycle arrest; 2-ME may be a potentially adjunctive anticancer drug useful to treat myelodysplastic syndrome.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Estradiol ; analogs & derivatives ; pharmacology ; Humans ; L-Lactate Dehydrogenase ; metabolism ; Myelodysplastic Syndromes ; pathology

OBJECTIVETo discuss the pathological characteristics of cervical spinal fracture complicating ankylosing spondylitis (AS) and the effect of combined anterior and posterior operation.

METHODSEighteen AS patients with traumatic cervical fracture-dislocation were treated operatively from January 2000 to January 2006. The symptom duration of AS was 14.5 years on average. Three cases had undergone osteotomy in lumbar spine. There were 4 cases of Grade A, 3 cases of Grade B, 9 cases of Grade C and 2 cases of Grade D according to Frankel's score. There were 15 cases of Grade III dislocation and 3 cases of Grade II. All patients underwent surgical procedures by combined anterior and posterior approach.

RESULTSThere were 4 anterior-posterior procedures, 8 anterior-posterior-anterior procedures and 6 posterior-anterior procedures. Seven patients had one stage operation and 11 cases underwent two stage surgeries. There was certain extent of neurological improvement in 14 incomplete paraplegic patients, but no improvement in 4 complete paraplegic patients. The follow-up period was 21.2 months on average and the time for bone fusion was 3.6 months. There were 4 complications during operation and a long-term complication in follow-up.

CONCLUSIONSThe study suggests that anterior combined with posterior approach makes the spine stable and relieves the pressure immediately. It is a reasonable surgical strategy for treatment of cervical spinal fracture-dislocation with AS.

Adult ; Cervical Vertebrae ; injuries ; Humans ; Joint Dislocations ; surgery ; Male ; Middle Aged ; Postoperative Complications ; etiology ; Retrospective Studies ; Spinal Fractures ; surgery ; Spondylitis, Ankylosing ; complications