BACKGROUND: Tissue-engineered materials containing osteoblasts have been widely used in the treatment of orthopedic diseases. How to improve hypoxic tolerance and the biological behavior of osteoblasts under hypoxic conditions is an important goal of the research on tissue engineering.OBJECTIVE: To explore the effect of M2 macrophage supernatant and Eucommia flavonoids on the biological behaviorof osteoblasts under hypoxia to pave ways for tissue engineering. METHODS:After MC3T3E1 cells were resuscitated, and M2 macrophages were isolated, induced and cultured, all cells were divided into control, M2 macrophage, Eucommia flavonoids and combination groups. The control group received no intervention. Cells in the M2 macrophage, Eucommia flavonoids and combination groups were exposed to M2 macrophages or/and 100 mg/L Eucommia flavonoids, respectively. RESULTS AND CONCLUSION:The cell viability in the M2 macrophage, Eucommia flavonoids and combination groups was significantly higher than that in the control group. The percentage of cells in the S and G2/M phases in the combination group was significantly higher than that in the control group. The relative mRNA expression levels of Runx2, collagen type Ⅰ and alkaline phosphatase, alkaline phosphatase activity and relative concentration of calcium ions in the combination group were significantly higher than those in the other three groups. To conclude, M2 macrophage supernatant combined with Eucommia flavonoids can promote osteoblast proliferation and differentiation under hypoxic conditions, maintain normal mineralization and osteogenesis of osteoblasts, and improve the tolerance ability of osteoblasts to hypoxia.

Objective To investigate the morphological alteration and the protein expression of caspase -3 in long -term denervated posterior cricoarytenoid muscles (PCAMs)in order to find the appropriate time point of rein‐neration in long -term denervated PCAMs .Methods A total of 45 patients with vocal paralysis were recruited and devided into 3 groups ,the 3~6 months denervation group ,the 6~12 months denervation group ,the 1~2 year denervation group .12 adults served as control group .The morphological alteration was evaluated using HE staining and the change in expression of caspase -3 ,an apoptosis related factor ,were observed using immunehistochemistry stain and western blot .Results With elongation of denervation time ,there were increased denaturation in the mus‐cle fibers .The nucleus moved inside and some of them concentrated .Caspase-3 showed weak staining in innerva‐ted ,however ,by 3-6 months of muscle denervation there was a significant accumulation of caspase -3 protein in myofibers ,6~12 months and 1~2 years of denervation ,expression of caspase -3 protein in myofibers was de‐creased significantly .In western blot ,the change in protein expression of caspase -3 was observed an 21-fold (P< 0 .01) increase from 3~6 months denervated muscles to innervated muscles ,11-fold (P< 0 .01) increase from 6~12 months denervated muscles to innervated muscles ,3~fold (P< 0 .01) increase from 1~2 year denervated muscles to innervated muscles .Conclusion The morphological alterations and changes in expression of caspase -3 indicated there was a high amplitude of apoptosis in denervated posterior cricoarytenoid muscles within 1 year .

Doxorubicin is widely used in chemotherapy of breast cancer and many other cancers.How ever,doxorubicin resistance restricts its use to some extent.The mechanisms of doxorubicin resistance may involve transport protein,apoptosis protein,DNA repair function,enzyme and other factors.Researches about reversing doxorubicin resistance are ongoing and provide some effective therapeutic regimens to overcome doxorubicin resistance clinically.But the specific mechanism of doxorubicin resistance is yet to be elucidated.It is expected that more reasonable and effective counter-measures will be put forward to improve the theraputical effect of doxorubicin.

Objective To study the effect of ursolic acid(UA) on apoptosis of the ovarian cancer cell line SKOV3 and its action mechanisms. Methods The SKOV3 cells were treated with UA at different concentration. The proliferation inhibition was examined by MTT assay. Morphological examination and flow cytometry were used to detect apoptosis. RT-PCR and immunocytochemical method were used to detect the expression of wild type p53,an apoptosis related genes. The semi-quantification of protein expression was analyzed by pathological image-analysis. Results UA inhibited the proliferation of SKOV3 cells strongly. Apoptosis of SKOV3 cells were induced by UA treatment. The morphology of SKOV3 showed changes such as chromatin aggregation, nuclear and cytoplasmic condensation and apoptotie bodies appearance. UA caused significant G0/G1 arrest with a concomitant decrease of cell population in S and G2/M phases. Apoptotic cells were found by flow cytometry. The maximal early apoptotic rate was (28.7±2.4)% when exposed to 40 μmol/L for 48 h. The expression of p53 gene and protein was enhanced. All these effects were in a dose and time-dependent manner Conclusion UA induced apoptotic processes in the ovarian cancer SKOV3 cell line were the mechanism involving p53 pathway.

In modern war, psychotropic drugs, including central stimulants and central depressants, have profound influence on the mental states of soldiers. This paper reviews the pharmacokinetics, pharmacological mechanisms and efficacy of psychotropic drugs: and also introduces the application of psychotropic drugs for sleep regulation and anti-fatigue in aircrew members during Iraqi war, South Atlantic Campaign, Libyan Air Strike and Gulf War. Suggestions are given on proper use of psychotropic drugs in wartime.

Objective: To study the morphological changes of posterior cricoarytenoid muscles (PCM) in patients with longterm denervation of recurrent laryngeal nerve (RLN), so as to provide theoretical evidence for repair of recurrent laryngeal nerve at advanced clinical stage. Methods: Thirty-eight patients with damaged RLN were divided into 4 groups according to the duration of their RLN damage: 6-12 months group (n = 12), 1-2 years group (n = 10), 2-3 years group (n = 8), and over 3 years group (n = 8). Twelve subjects with normal PCM served as control. Trichrome Masson staining and imaging analyzing system were used to quantitatively analyze the transverse section areas of myofibers, collagen fiber and connective tissues. SDH and AchE staining and cell counting method were used to analyze changes of two kinds of myofibers and motor end plate numbers at different times after denervation of recurrent laryngeal nerve. Results: The transverse areas of myofibers gradually decreased and those of collagen fibers gradually increased with the prolongation of denervation; the difference was significant between different groups (P<0.01). The ratio of transverse area of myofiber to that of collagen fibers reached the lowest level 0.5-2 years after denervation. The fibrosis of muscle obviously slowed down 2 years after denervation. The transverse section of 48% of myofibers remained 3 years after denervation, Long-term denervation resulted in the changes of muscle fiber types: the ratio of red muscles was increased and the ratio of white muscles was decreased after denervation; the difference was significant between different groups (P < 0.01). The number of motor end plate decreased with the prolongation of denervation and disappeared after 1 year. Conclusion: The morphological alteration in long-term denervation PCM indicates the worst myofibrosis occurs within 2 years of denervation, but 48%of myofibers remain 3 years after denervation. The type alteration of denervated muscles may decrease the apoptosis of skeletal muscle. The structure of myoceptors disappears within 1 year of denervation. Our experiment indicates that there is a morphological basis for regaining total or partial muscle function by nerve repair after long-term denervation.

Objective: To investigate the change in myogenin expression at different time in long-term denervated human posterior cricoarytenoid muscles (PCAMs), so as to provide a theoretical basis for timing of reinnervation. Methods: Thirty-eight specimens of denervated human PCAMs were divided into 4 groups according to the period of denervation: 6-12 months denervation group(n= 12), 1-2 years denervation group (n = 10), 2-3 years denervation group (n = 8), and over 3 years denervation group(n=8). Another 12 specimens of normal PCAMs served as control. The patients in all groups were age- and sex-matched. The expression of myogenin protein and mRNA was studied using immunofluorescence staining and real-time PCR analysis, respectively. Results: Immunofluorescence staining showed that the positive myogenin expression was mainly found in the myonuclei of PCAMs with a denervation period less than 3 years; no positive staining was found in the myonuclei of control group. The expression of myogenin in myonuclei and the ratio of positive cells were up-regulated in the 6-12 month denervation group compared with those in the control group; the expression and the ratio peaked in 1-2 years denervation group and decreased again in the 2-3 years denervation group, but was still significantly higher than those of the control group (all P< 0.01). There was hardly any expression of myogenin 3 years after denervation. Results of RT-PCR showed no myogenin mRNA expression in the control group; the expression in 1-2 years, 2-3 years, and more than 3 years denervation groups were 4 times, 64 times, and half that of the 6-12 months denervation group, respectively (all P<0.05). Conclusion: It is indicated that there is a potential for muscle regeneration within 3 years of denervation.

Objective: To investigate the in vivo absorption kinetics of caffeine at different intestine segments in rats. Methods: The rat intestine was cannulated for in situ recirculation. The absorption kinetics of caffeine, including the absorption segments of intestine, drug concentration and pH value, were investigated in all the rats under light anesthesia. Ultraviolet light spectrometry and high-performance liquid chromatography were used to determine the concentrations of phenol red and caffeine, respectively. Results: The absorption rate constants (Ka) of caffeine in the small intestine at the concentrations of 2.5, 5, 10 and 25 mg · L-1 were 0.252, 0.247, 0.304 and 0.232 h-1, respectively. The Ka values at pH of 7.8, 6.8, and 5.4 were 0.267, 0.274 and 0.247 h-1, respectively. The Ka values in the duodenum, jejunum, ileum, and colon were 0.112, 0.099, 0.095 and 0.069 5 h-1 respectively. Conclusion: Concentration and pH value have no obvious effect on the absorption kinetics of caffeine. Caffeine can be better absorbed in the duodenum,jejunum and ileum than in the colon. The absorption of caffeine in the intestine is a first-order process through passive diffusion mechanism.

OBJECTIVE: To evaluate the effect of Eucommiae ulmoides on the microstructural and ultrastructural change of femur and lumber vertebrae in ovariectomized rats.METHODS: 80 4-month-old female SD rats were randomized to 4 groups: sham operation group,model group,alfacalcidol group,E.ulmoides group.In the sham operation group,the abdominal cavities of the rats were cut open to expose ovaries before being sutured.However,the rats in the other 3 groups were ovariectomized bilaterally.At 10 days after operation the rats were administered intragastrically with corresponding drug.After 3 months,femur and lumber vertebrae were dissected and fixed in 4% glutaral for 3 days,then decalcified in 4% EDTA for 4 weeks,and then the structure of femur and lumber vertebrae was observed under light microscope and transmission electron microscope(TEM).RESULTS: Observed under light microscope,the femur and lumber vertebrae were full of fat in the ovariectomized rats and in which the density of bone trabecula decreased significantly;however,in alfacalcidol-treated rats and E.ulmoides-treated rats,the amount of fat decreased markedly while the density and thickness of bone trabecula increased.Observed under TEM,in the ovariectomized rats,rough endoplasmic reticulum(RER),cytolysosome and mitochondrial were plentiful inside osteoclasts in femur and lumber vertebrae,partial osteoblasts atrophied,the amount of RER and other cellular organ decreased significantly and apoptosis appeared for partial cells;however,in alfacalcidol-treated group and E.ulmoicdes-treated group,the above changes improved significantly.CONCLUSION:E.ulmoides has remarkable beneficial effect on osteoporosis in ovariectomized rats.The mechanism might be attributed to its inhibition on the synthesis of fat and the activity of osteoclasts,promotion on the activity of osteoblasts and inhibition on the apoptosis.

Humans ; Korea ; Medicine, Chinese Traditional