Previous methods used for nuclear transplantation were further investigated to develop a method that was both easy to carryout and did not require any special apparatus, such as Piezoimpact or Spindle-View. Following the puncture of zona pellucida with two holes by injection pipette that contained donor nuclei or cells, the injection pipette was pulled back to the perivitelline space while the negative pressure was increased in the holding pipette until the polar body and karyoplasm were wiped off completely. Then a reconstructed embryo was completed by the direct injection of the donor nucleus or cell without pulling out the injection pipette. 200 oocytes were manipulated using this method and it cost about 40 seconds with nucleus injection and about 30 seconds with cell injection to complete a reconstructed embryo. The success rates were 62.6% and 86. 0%, respectively, and enucleation rate was about 73.3% validated by Hoechst 33342. Using this method, the nucleus was completely eliminated and another was injected using the microscope and micromanipulator. Moreover, the efficiency of nuclear transplantation and survival rate of reconstructed embryos were greatly improved. Furthermore, it is very easy to manipulate and popularize in practice.
Animals ; Cell Culture Techniques ; methods ; Cell Nucleus ; metabolism ; Cells, Cultured ; Cloning, Organism ; methods ; Embryo, Mammalian ; cytology ; metabolism ; Embryonic Development ; Female ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Inbred DBA ; Mice, Inbred Strains ; Nuclear Transfer Techniques ; Oocytes ; cytology ; metabolism ; Zona Pellucida ; metabolism

This essay is to present an improvement on the concentration assay for hemihydrate gypsum in plaster of Paris bandage-Viscose form.
Calcium Sulfate ; analysis ; Casts, Surgical ; Titrimetry ; methods

OBJECTIVETo analyze the results and the prognosis of local excision of lower rectal cancer, and investigate the proper indication of local excision for cure purpose.

METHODSThe clinicopathological data of 76 patients with lower rectal cancer treated from February 1985 to October 2007 were analyzed.

RESULTSSixty-nine patients received transanal excision, 6 cases received trans-sacral excision and 1 case received trans-vaginal excision. Among the cases, 48 cases were ranged as T1 phase, 25 cases as T2, 3 cases as T3. The operation complication rate was 7.9%, and the 30-day mortality rate post operation was 0. The local recurrence rate was 22.4% and the overall 5 year survival rate was 84.5%. The local recurrence was significantly related with T stage and resection margin status. The survival was significantly related with mucinous adenocarcinoma, resection margin status and lymphovascular invasion; and the resection margin and lymphovascular invasion was the independent factors affecting survival.

CONCLUSIONLocal excision was safe in early stage lower rectal cancer. Careful patient selection is the key of the operation. The proper indication of local excision is T1-2 without high risk factors (high histopathological grade, presence of lymphatic or vascular invasion, mucinous adenocarcinoma); and the cases with T2 phase tumor should undergo chemoradiotherapy after local excision.

Adult ; Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Prognosis ; Rectal Neoplasms ; pathology ; surgery ; Retrospective Studies ; Survival Analysis ; Treatment Outcome

OBJECTIVETo study the correlation between the expression of Egr-1 and NF-kappaB and the up-regulation of TNF-alpha and TGF-beta1 in macrophages after stimulation by silica in-vitro.

METHODSMacrophages were treated with antibodies against Egr-1 and NF-kappaB and antisense oligonucleotides. The level of TNF-alpha protein in the cell supernatant was then measured using enzyme-linked immunoadsorbent assay (ELISA). The expression of TGF-beta1 protein was detected by immunocytochemistry. The expression of TNF-alpha and TGF-beta1 mRNAs was also monitored by reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTSCompared with silica-stimulated macrophages untreated with antibodies, the cells treated with 10 micro g/ml of Egr-1 or NF-kappaB antibodies were associated with reduced expression of TNF-alpha and TGF-beta1 proteins and mRNAs (P < 0.05). Compared with silica-stimulated untransfected group, the antisense group was associated with obvious reduction in the expression of TNF-alpha and TGF-beta1 proteins and mRNAs (P < 0.05).

CONCLUSIONThe expression of TNF-alpha and TGF-beta1 mRNAs and proteins are associated with activation of Egr-1 and NF-kappaB in macrophages, after stimulation by silica. It is possible that the corresponding antibodies and antisense oligonucleotides may become a potential therapeutic tool in the management of silicosis in the future.

Animals ; Antibodies ; immunology ; Cells, Cultured ; DNA-Binding Proteins ; genetics ; immunology ; Early Growth Response Protein 1 ; Immediate-Early Proteins ; genetics ; immunology ; Macrophages ; cytology ; metabolism ; Mice ; NF-kappa B ; genetics ; immunology ; Oligonucleotides, Antisense ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Silicon Dioxide ; pharmacology ; Silicosis ; etiology ; Transcription Factors ; genetics ; immunology ; Transforming Growth Factor beta ; biosynthesis ; genetics ; Transforming Growth Factor beta1 ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics

OBJECTIVETo establish a method for real-time recording the oxygen consumption of mice under normobaric hypoxia.

METHODSThe experimental apparatus was made up of animal container, filling water control system, electronic balance, hose, a computer with weight recording software, etc. The working principle was that the oxygen consumed by animal was replaced by water filling which was controlled by the pneumatic and hydraulic actuator. The water was weighted by an electronic balance and the weight signal was recorded into excel file at the same time. The accuracy and precision of the apparatus were detected by a 10 ml syringe. The oxygen consumption characteristics of 6 acute repetitive hypoxia mice and 6 normal mice were observed.

RESULTSThe P value for the paired t test was 1 and the CV value was 4%. The survival time and total oxygen consumption of acute repetitive hypoxia mice were both significantly increased compared to normal mice (P < 0.05), which were (58.8 +/- 6.8) min and (46.0 +/- 8.7) min respectively for the survival time and (85.1 +/- 8.5) ml and (73.6 +/- 5.4) ml respectively for total oxygen consumption.

CONCLUSIONThe hypoxia tolerance of the acute repetitive hypoxia mice can significantly improved by taking more oxygen in the animal cabin. The accuracy and precision of the apparatus are high and it can be used for the determination of oxygen consumption in hypoxia research.

Animals ; Hypoxia ; physiopathology ; Mice ; Monitoring, Physiologic ; instrumentation ; Oxygen Consumption ; physiology

BACKGROUNDAnorectal malignant melanoma was a rare disease with extremely poor prognosis. The aim of this study was to explore the clinical characteristic, diagnosis and treatment strategies of anorectal malignant melanoma.

METHODSThe data of 57 patients with anorectal malignant melanoma was collected and retrospectively analyzed.

RESULTSRectal bleeding and anal mass were found to be common symptoms of anorectal malignant melanoma. The preoperative diagnosis rate of anorectal malignant melanoma was 48.6%. The overall 3-year and 5-year survival rate was 38.0% and 21.3% respectively. The 3-year survival rates of stage I and II patients were 63.0% and 16.7% respectively (P = 0.000), and the 5-year survival rates were 33.3% and 11.1% (P = 0.001), which both had significant statistic differences. The 3-year survival rate of patients undergone abdmoninoperineal resection and patients undergone wide local excision were 36.7% and 53.0% respectively (P = 0.280), while the 5-year survival rate were 24.1% and 23.1% (P = 0.642), which both had no significant statistic differences.

CONCLUSIONSThis study identified no survival advantage to abdominoperineal resection in treatment of anorectal malignant melanoma, and we propose that wide local excision could be considered as the initial treatment of choice.

Adult ; Aged ; Aged, 80 and over ; Anus Neoplasms ; diagnosis ; mortality ; pathology ; surgery ; Female ; Humans ; Male ; Melanoma ; diagnosis ; mortality ; pathology ; surgery ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Survival Rate

OBJECTIVETo find out the vector ability and function of Nosopsyllus wualis leizhouensis in the transmitting plague.

METHODSIn T: 19 degrees C +/- 1 degrees C, RH: 85% +/- 5%, data regarding the vector ability as cluster spreading, single flea spreading, single flea transmitting plague to single animal, formative bacterial embolus and infection fleas life-span through experiments was gathered.

RESULTSThe rate of infection on fleas was 94.64%, with 100% transmission rate of colony to spread, and 30% from single flea spreading to single animal. In the experiment of single flea transmission, all of the 388 rattus loseas were bitten by the fleas with bacterial, but only 9 animals were characteristically infected with the transmission potential, vector efficiency, survival potential of embolus, vector index as 0.360, 0.257, 0.868 and 0.223 respectively. The mean survive days of infected flea feed with blood were 17.58 (1 - 58), and the mean survive days of hunger infected flea were 7.25 (1 - 16). Formative bacterial embolus days were 8.80 (2 - 16) and the rate of embolus flea was 78.12%.

CONCLUSIONNosopsyllus wualis leizhouensis could serve as vector and important in the mode of plague transmittion.

Animals ; Female ; Insect Vectors ; microbiology ; Male ; Plague ; transmission ; Rats ; Siphonaptera ; microbiology

OBJECTIVETo investigate the effects of SiO(2) on the expression of alpha-smooth muscle actin (alpha-SMA) in human lung fibroblasts in vitro and vivo.

METHODSThe experimental group comprised 32 rats while 32 rats were included in the control. In vivo, the expression of alpha-SMA in lung tissues of rats exposed to SiO(2), the supernate of RAW264.7 cells, SiO(2) and the growth factor beta(1) (TGF-beta(1)) were investigated, respectively.

RESULTS(1) alpha-SMA positive myofibroblasts appeared in the lung tissues of the 28th day groups exposed to SiO(2). (2) The expression of alpha-SMA in HLF-02 cells was unregulated by TGF-beta(1) and supernate of RAW264.7 cells exposed to SiO(2). (3) The expression of alpha-SMA in HLF-02 cells was not induced by SiO(2).

CONCLUSIONMyofibroblasts related to silicosis, and the appearance of myofibroblasts (in vitro) are independent on direct stimulation by SiO(2), but related to the mediator (TGF-beta(1)) secreted by SiO(2) stimulated macrophages.

Actins ; biosynthesis ; genetics ; Animals ; Cells, Cultured ; Fibroblasts ; drug effects ; metabolism ; Lung ; cytology ; metabolism ; Macrophages, Peritoneal ; drug effects ; Rats ; Rats, Sprague-Dawley ; Silicon Dioxide ; pharmacology ; Silicosis ; metabolism ; pathology ; Transforming Growth Factor beta1 ; pharmacology

OBJECTIVETo explore the clinical value of magnetic resonance (MR) imaging in the diagnosis of the complications after breast augmentation with polyacrylamide hydrogel injection.

METHODSMRI was performed in 16 patients who had breast augmentation with polyacrylamide hydrogel injection. The results of magnetic resonance imaging were compared with the clinical symptoms.

RESULTSMRI examinations of the 16 patients showed that (1) 4 breasts in 2 patients were comparatively natural; (2) a large lacteal cyst was detected in one patients in lactation; (3) breast asymmetry occurred in 3 patients (6 sides); (4) in 10 patients (20 sides), the injected gel was in irregular form. The results of MRI were in accord with the clinical symptoms.

CONCLUSIONMRI is of great diagnostic value for the complications after breast augmentation with polyacrylamide hydrogel injection.

Acrylic Resins ; administration & dosage ; Adult ; Breast ; pathology ; Female ; Humans ; Injections ; Magnetic Resonance Imaging ; Mammaplasty ; adverse effects ; methods ; Postoperative Care ; Postoperative Complications ; diagnosis ; etiology ; Sensitivity and Specificity

OBJECTIVETo investigate the effect of total astragalosides (TA) on proliferation and apoptosis in human leukemia NB4 cells in vitro.

METHODSThe NB4 cells were treated with TA at different concentrations for 48 h in culture. Growth inhibition rates were measured by CCK-8 method. Flow cytometry was used to explore the cell apoptosis and the activity of NF-κB and Akt during apoptosis.

RESULTSTA at different concentrations (200, 400, 600, 800 mg/L) inhibited proliferation of NB4 cells in a dose-dependent manner (P < 0.05), and the inhibitory rates of TA on NB4 cells were (14.54 ± 3.20)%, (24.79 ± 3.98)%, (57.28 ± 4.71)% and (88.28 ± 4.65)%, respectively. In terms of the induction of apoptosis, there was a significant difference between the TA group and blank control [(1.80 ± 1.24)%, P < 0.05]. At TA doses of 200, 400 and 600 mg/L, the apoptotic rates of NB4 cells were (10.03 ± 3.31)%, (14.87 ± 3.65)%, (23.45 ± 1.90)%, respectively. Besides, TA induced apoptosis of NB4 cells in a dose-dependent manner in the groups of 200 mg/L, 400 mg/L, 600 mg/L (P < 0.05). But there was no significant difference in apoptotic rates between the groups of 800 mg/L and 600 mg/L [(23.45 ± 1.90)%, P > 0.05]. In the group of 800 mg/L, the necrotic cells increased highly and the necrotic rate reached (45.65 ± 3.16)%. After TA treatment of NB4 cells at different concentrations (200, 400, 600 mg/L), the expression of NF-κB protein was significantly decreased compared with that of the blank control (9.79 ± 0.95, P < 0.05), while Akt protein was not significantly decreased (P > 0.05).

CONCLUSIONTA can inhibit the growth of NB4 cells and induce apoptosis in NB4 cells through an Akt-independent NF-κB signaling pathway.

Antineoplastic Agents, Phytogenic ; administration & dosage ; isolation & purification ; pharmacology ; Apoptosis ; drug effects ; Astragalus membranaceus ; chemistry ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Humans ; Leukemia, Promyelocytic, Acute ; metabolism ; pathology ; NF-kappa B ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Saponins ; administration & dosage ; isolation & purification ; pharmacology