BACKGROUND: Vitrification is a comparatively new technology which applies high concentration cryoprotectant and rapid refrigeration. By the method, the cells were quickly frozen and to avoid damage by ice crystals inside and outside. OBJECTIVE: To compare the effect of four cryoprotectants on morphology and function of ovarian tissue in rats after vitrification. METHODS: The rats were randomly assigned into six groups with 6 rats for each: DMSO + EG, DMSO + EG + sucrose, DMSC +EG + sucrose + acetamide, EG + sucrose + acetamide, ovariectomized, and normal control groups. The ovarian tissues of four freezing groups were treated with the corresponding cryoprotectants, the vitrified ovarian tissues were then resected but not frozen and transplanted; otherwise, tissues were not treated with any treatment in the normal control group. Two weeks after freezing, the tissues were thawed and heterotopic-transplanted into femoribus intemus of hind limb. At 30 days after implantation, vaginal epithelial cells and estrus cycle were observed, while after three months, blood were collected to detect the level of estradiol (E2) and the ovarian tissues were reclaimed to analyze their morphological changes. RESULTS AND CONCLUSION: All ovarian tissues were damaged after cryoprersarvation in four freezing groups. The rates ot healthy primordial follicles were 67.9%, 71.6%, 80.5%, and 59.4%, respectively, while healthy primary follicles were 41.6%, 52.3%, 55.9%, and 36.7%, respectively. In all freezing groups, the rate of the healthy follicles in DMSO + EG + sucrose + acetamide group was higher than DMSO + EG group and EG + sucrose + acetamide group (P < 0.05). No significant difference was found in the proportion of follicles at different development stages among four groups. The typical secondary follicle was not found in four groups. Damaged ovotid showed oocyte pyknosis and vacuolation in cytoplasmic area. There was not typical cell type of all freezing groups. Ovarian autografting gained visible vascularity from surrounding tissue that connected ovarian tissue to form net. There was a lot of blood capillary in transplanted ovarian tissues and clumped primordial follicles in cortical substance. The rates of primary follicles and secondary follicles were lower than primordial follicles. The level of serum estradiol was obviously decreased compared with normal control group (P < 0.01). There was significant difference between DMSO + EG + sucrose + acetamide group and other three freezing groups (P < 0.05). Four kinds of freezing methods have poor effects on different stages of follicles and the structure of ovariarn tissue. DMSO + EG + sucrose + acetamide group is an optimal protocol for cryoprerserving ovarian tissue. Freezing methods still need to explore further because the rats had not appeared disciplinary estrus cycle after ovarian autoqrafting.

BACKGROUND: What deserves exploration is the relationship between menstrual cycle phase and athletic ability of female athletes, especially the mechanism of athletic performance influenced by different menstrual cycle phases.OBJECTIVE: To compare the hormone level of different menstrual cycle phases and the changes of athletic performance as well as its effect on athletic performance.DESIGN: Single sample and single factor analysis.SETTING: Zhejiang Insftute of Sport Science; College of Education,Zhejiang University; Department of Obstetrics and Gynecology, Zhejiang Provincial People's Hospital.PARTICIPANTS: Specially designed menses card were distributed to all the female athletes in the gig team and track & field team in Zhejiang Province in 2000. Consecutive 3-month recording was made to investigate the status of menstrual cycle phases of female athletes. Twenty-five athletes with regular menses were chosen as subjects, among whom 12 were specialized in gig racing with the average age of 19 years and 13 were specialized in track & field events with the average age of 18 years. They all had more than 3-year training experience, 6 days a week and 4-6 hours a day. They were healthy, had regular menstrual cycle, and had not taken any drug which influences the menstrual cycle.METHODS: Blood samples of the testees were collected at hour 8-9 AM at follicular phase (days 6-10) and luteal phase (days 19-23). Full-automatic chemiluminescence immunoassay equipment was used to measure the level of estradiol (E2), follicle-stimulating hormone (FSH), luteinizing hormone (LH), progesterone, and testosterone in serum. coefficient of variability in batch of reagent was ＜5%. Testees accomplished test of their athletic performance and the test of lactic acid 1 to 2 days after blood sample collection. Gig athletes performed 2000 m and 500 m full-strength pull with Concett Ⅱ boating ergometer while track & field athletes performed 100 m and 200 m full-speed running; the time needed was recorded. Gig athletes conducted 2000 m movements on the ergometer with the frequency of 26 times per minute. Track & field athletes carried out three times of 300-meter running with 90% of intensity with 3-minute intervals. Immediately after movements, the lactic acid value was measured following blood collection.Paired t test of the small sample study was used for significance detection.MAIN OUTCOME MEASURES: The level of E2, FSH, LH, proges-terone and blood lactic acid in serum; 2000 m and 500 m performance of gig athletes as well as 100 m and 200 m performance of track & field athletes were measured by the ergometer.RESULTS: Twenty-five athletes accomplished detection of all the indexes and entered the result analysis. The level of FSL, progesterone and testosterone of gig athletes was higher at luteal phase than at follicular phase;however, E2 and LH level did not significantly differ. The level of progesterone and testosterone of track & field athletes was higher at luteal phase than at follicular phase; however, E2, FSH and LH level did not significantly differ. 500 m performance of gig athletes detected by the ergometer, and 100 m and 200 m full-speed running performance of track & field athletes at luteal phase were superior to those at follicular phase, but no significant difference was found in 2000 m performance of gig athletes. When the same loading was accomplished at follicular phase and luteal phase, the lactic acid value at luteal phase was significantly lower than that at follicular phase.CONCLUSION: Tested athletes at luteal phase have good functional status, which may be related to the changes of menstrual cycle hormone level and synergistic effect of various hormones under thes state of athletic movements. It suggests that sufficient attention should be paid to the menstrual cycle of female athletes.

Objective: To study the phenotype characteristic of mandibular condylar chondrocytes (MCCs) in vitro. Methods: MCCs of two-week-old New Zealand rabbits were isolated with enzyme digestion and cultured in DMEM. Phase contrast microscopy, image analysis system, MTT assay and immunocytochemistry method were used to compare MCCs of different passages in cellular shape and size, growth curve and expression of type I and type II collagen. Results: The majority of MCCs in earlier passages (1～3 passage) were polygonal, while more fusiform and spindle-shaped cells were found after 5～6 passages. MCCs in earlier passages were smaller than those in sixth passages. The proliferation rate of MCCs decreased with passaging. There were more type II collagen positive cells in 1 ～ 3 passages, while more type I collagen positive cells in 7th passage. Conclusion: The changing characters of MCCs in vitro such as cellular shape and size and expression of type I and II collagen are similar to those of MCCs in vivo from proliferating zone to hypertrophic zone. MCCs in later passages may be with dedifferentiation.

Objective To explore the effect of histamine in paraventricular nucleus (PVN) on the development of neurogenic pulmonary edema (NPE).Methods NPE models of rabbits were made and H 1 receptor antagonist (chlorpheniramine, H 1 group), H 2 receptor antagonist (Cimetidine, H 2 group) or normal sodium (NS group) were injected into PVN of rabbits utilizing the technique of stereotaxis. The degree of pulmonary edema was observed, the content of histamine in PVN was measured by high performance liquid chromatogram, and the ration of lung-water and A/P value were calculated.Results (1) Compared with control group and sham operation group, the content of histamine of PVN in NPE group obviously increased ( P

[Objective]To evaluate tho effects of Bone Mesenchymal Stem Cells(BMSCs) on healing of bone-tendon junction in a patellar tendon complex.[Method]The BMSCs of rabbits were harvested by bone marrow aspiration. Adherent cells were selected as BMSCs after the whole marrow was cultured.Standard partial patellectomy was conducted in 36 18-week-old rabbits divided into 2 groups. The experimental group was planted into by the composite of BMSCs and Pluronic F-127 while the control group was only operated without BMSCs planting into.The patellar-patellar tendon complexes were harvested at weeks 6,12,18 postoperatively for radiographic, gross and histological evaluations.[Result]Radiographic measurements showed significantly more newly formed bone at the patellar tendon-patellar healing junction in the experimental group compared with that of the controls at weeks 6,12,18(P

AIM: To establish the quality standard for Yinchai Granules (Caulis Lon icerae, Radix Bupleuri, Folium Eriobotryae, etc). METHODS: TLC was used to identify Radix Bupleuri, Folium Eriobotrya e and HPLC to determinate the chlorogenic acid content was determined by HPLC. RESULTS: The linear range of chlorogenic acid was 23.64～378.24ng, r=0.9999, respectively. The average recovery was 99.14%. CONCLUSION: The method is simple. The result is accurate and the reproducibility is good and can be used as the quality control of Yin chai Granules.

Patients on Porcelain-fused-to-metal restorations of vital teeth are increasingly year by year, this article mainly discusses the psychological characteristics of adult on porcelain-fused-to-metal restorations of vital teeth on the view of medical ethics.It is helpful to lighten patients’burden and complete treatment successfully by improving the qualities of dentists and giving patients psychological guidance.

Objective:To establish a method for the determination of related substances in enoxacin gluconate injection by HPLC with gradient elution. Methods:HPLC was performed on a DiamonsilTM C18 column (250 mm × 4. 6 mm, 5 μm), the mobile phase A was 0. 025 mol·L-1 phosphonic acid solution (adjusting pH to 3. 0 with triethylamine)-methanol- acetonitrile (80∶10∶10) and the phase B was 0. 025 mol·L-1 phosphonic acid solution (adjusting pH to 3. 0 with triethylamine)-methanol-acetonitrile (350∶325∶325) with gradient elution at a flow rate of 1. 1 ml·min-1 , the detection wavelength was 269 nm and 284 nm, the injection volume was 20μl, and the column temperature was 40℃. Results:Under the HPLC conditions, the samples had good stability and separation. The calibration curve was linear within the concentration range of 19. 80 ng·ml-1-19. 80 μg·ml-1(r=0. 999 9) for 5-hydroxymethylfur-fural with the detection limit of 0. 18ng, and the average recovery was 99. 68% with RSD of 0. 12% (n=9). Conclusion:The method is accurate, sensitive, specific and reproducible, and can be used in the determination of related substances in enoxacin gluconate in-jection.