The effects of different carbon sources added to aerobic medium on cell growth,enzyme activity and metabolite distribution was investigated in dual-phase fermentations.The results showed that both cell density and enzyme activity were enhanced by adding 4mmol/L glucose or 12,54,80mmol/L sodium acetate to aerobic medium,respectively.Then centrifugated cells that had grown aerobically in different conditions were transferred to anaerobic fermentation,the enzyme activity and metabolite distribution changed.To analyze the anaerobic enzyme activity and metabolite distribution,it was concluded that during the anaerobic fermentation of Escherichia coli NZN111 overexpressed malic enzyme,PEP carboxykinase(PCK)was the key enzyme of succinate production,pyruvate kinase(PYK)was associated to the accumulation of by-product pyruvate,and isocitrate lyase(ICL)have some influence on the concentration of succinate.The yield of succinate in anaerobic stage could reach 89.0%,which was 16.6% higher than that of the control by adding 80mmol/L sodium acetate as carbon source in aerobic medium.

BACKGROUND: Fat embolism syndrome is a commonly seen severe complication in the field of opthopaedics in clinic. It frequently occurs after long bone fracture and hip-knee joint replacement. However, its etiological factors and pathogenesis are not identified.OBJECTIVE: To sum up the onset influencing factors, pathogenesis and therapeutic methods of long bone fracture and hip-knee joint replacement accompanied with fat embolism syndrome.RETRIEVE STRATEGY: Using the terms "fat embolism syndrome, pathogenesis, treatment, prevention, diagnosis", we retrieved PubMed database to identify studies published in the English language. Fifty-five literatures were searched.Meanwhile, we searched the medical information network of Shanghai Jiao Tong University with the same terms in the Chinese language. Inclusive criteria: studies, which can reflect the diagnosis and treatment as well as pathogenesis of fat embolism syndrome. Exclusive criteria: repetitive studies.LITERATURE EVALUATION: The involved 38 literatures are all about the diagnosis and treatment as well as pathogenesis of fat embolism syndrome, among which, 6 were review and the others were clinical or basic studies.DATA SYNTHESIS: ① The influencing factors of fat embolism syndrome included trauma factor, operation factor and other factors. ② The pathogenesis of fat embolism syndrome involved mechanical obstruction theory, biochemical theory,condensation theory and inflammatory reaction theory. ③ The treatments of fat embolism syndrome include respiration supporting, glucocorticoid application, protecting brain function, drug treatment, heat shock treatment and so on.CONCLUSION: Study on bone fracture and joint replacement accompanied with fat embolism syndrome can provide evidence for the diagnosis and treatment of this syndrome.

Objective To quantitatively observe the actual levels of 4977 bp and 3895 bp mitochon-drial DNA (mtDNA) deletion in healthy human skin of different age, and to explore their relationship with intrinsic aging and photoaging. Methods Seventy-one samples of skin tissue were obtained from healthy volunteers, including 40 samples from UV-protected areas (buttock, thigh, waist or abdomen) and 31 from UV-exposed areas (neck, back of hands, forehead or face). Nuclear and mitochundrial DNA was extracted from these samples. Conventional PCR was performed to detect the incidence of 4977 bp and 3895 bp mtDNA deletion. Then, SYBR Green real-time PCR was applied to quantitatively analyze the target mutations in posi-tive samples. Results Conventional PCR showed that the incidence of both 4977 bp deletion and 3895 bp deletion increased with age. For example, the incidence of 4977 bp deletion and 3895 bp deletion accounted for 47.5% (19/40) and 30% (12/40), respectively in samples from volunteers older than 40 years, signifi-cantly higher than that in those from volunteers younger than 40 years (X2 = 4.673, 6.118, respectively, both P < 0.05). The total incidence of 4977 bp deletion and 3895 bp deletion in UV-exposed areas was 48.4% (15/31) and 32.3% (9/31), respectively, which did not differ from those in UV-protected areas. The results from real-time PCR revealed a positive correlation of the copy number of 4977 10p deletion and 3895 bp deletion with age (rg = 0.907, 0.845, respectively, both P < 0.05). When the UV-exposed area was compared with the UV-protected area, no significant difference was found in the copy number of 4977 bp deletion ( P = 0.264), whereas a higher level of 3895 bp deletion was noticed in UV-exposed area (P = 0.014). Conclu-sions The 4977 bp mtDNA deletion is primarily associated with chronological aging, and might serve as a biomarker for the process of chronological aging of skin. Deletion of 3895 bp mtDNA seems to be more sus-ceptible to ultraviolet radiation.

Objective To observe the immune regulatory effect of Marasmius androsaceus (MA) polysaccharide in normal mice. Methods The experimental mouse was injected with high,middle,and low dosages of marasmius androsaceus polysaccharide(5,10,20 mg?kg-1?d-1) in the abdomen (i.p.) ,and the comparison group was given the same volume of saline water i.p. once a day. After treatment for 14 consecutive days,the blood cell count instrument was used to measure the mice hemogram,the phagocytic function of peritoneal macrophage and the ability of carbon granular clearance were observed.,the ability of the spleen lymphocytes proliferation was tested by MTT,and the hemolysin production test was used to evaluate the influence of MA polysaccharide on specific humoral immunity in mice. Results Marasmius androsaceus polysaccharide at middle and high dosages can increase the number of lymphocytes and moncytes in normal reference,enhance the phagocytic function of peritoneal macrophage,promote the carbon granular clearance,accelerate the proliferation of T lymphocyte,enhance the specific humoral imnune function. Conclusion Marasmius androsaceus polysaccharide has certain potentiating effect on the phagocytic function and immunity regulation.

Objective To establish and characterize human colorectal cancer orthotopic implantation model in nude mice.Methods Established the human colorectal cancer orthotopic implantation model by surgery,characterize its biological characteristics by immunohistochemistry.Results Human colorectal cancer orthotopic implantation model was established in nude mice successfully,the proliferating cell nuclear antigen and Survivin were overexpressed in the models which were simulated the characteristics of human colorectal cancer perfectly.Conclusion This human colorectal cancer orthotopic implantaion model will provide an excellent animal model for us to study the development and metastasis mechanism of human colorectal cancer.

Objective To observe the aging,apoptosis,cell cycle arrest and oxidative stress in human skin fibroblast(HSF)induced by UVB,and to detect the expression profiles of p66Shc,a determinant of oxidative stress response and life span,in this process.Methods HSF cells were exposed to UVB at a subcytotoxic dosage twice a day for three days.The cells without exposure served as control.After another 24-hour culture,SA-β-Gal staining was performed to evaluate the senescence state of the cells,flow cytometry to observe cell apoptosis;cell cycle arrest was detected by serum starvation and flow cytometry:ELISA was applied to detect intracellular levels of superoxide dismutase(SOD)and malondialdehvde(MDA),and Western blotting to analyze the expression of p66Shc protein.Results The percentage of cells positive for SA-β-Gal staining increased from 0 to 98.3% after UVB radiation,which strongly suggested an aging state of HSF cells.The percentage of apoptotic cells increased from 0.96% to 37%.and 80.07% of the HSF cells were arrested in G0/G1 phase following the irradiation.Intracellular SOD activity decreased from(52.35±4.97)ng/g to(7.81±0.68)ng/g(P＜0.01).while intracellular MDA was found to increase from(3.52±0.34)ng/g to(33.91±3.20)ng/g(P＜0.05).The p66Shc protein was found to be weakly expressed in HSF in 24 hours following the exposure to UVB,and a stronger expression was noted 48 hours later.Conclusions HSF cells are induced into a state of senescence associated with oxidative stress after UVB irradiation,which may be applied as an in vitro model in aging research.The expression of p66Shc is increased in HSF during this process,and further studies are needed to explore the relation between p66Shc and oxidative stress as well as cellular aging.

Objective: To investigate the effect of chronic intermittent hypoxia (CIH) on liver injury and the expression of fractalkine in rats and explore its possible mechanism.
Methods: A CIH murine model was established to mimic the pathophysiology of obstructive sleep apnea-hypopnea syndrome (OSAHS) in humans. Thirty healthy male Spraque-Dawley rats were randomly assigned to 3 groups: a 5% CIH group, a 5% CIH+RH (removal of hypoxia) group and a control group ( 10 rats in each group). The 5% CIH and 5% CIH+RH groups were exposed to CIH for 3 weeks, 8 h/d, and the frequency of hypoxia was 20 times/h. The 5% CIH+RH group was then exposed to normal gaseous environment for another 3 weeks. After the experiment, liver sections were stained with hematoxylin-eosin (HE) and the liver pathology was observed. The expression of fractalkine in the liver tissues was detected by immunohistochemical method.
Results: 1) Compared with the control group, the hepatic steatosis and inflammatory activities in the 5% CIH and 5% CIH+RH groups were more severe (allP<0.01 ); compared with the 5% CIH group, the hepatic steatosis and inflammatory activity in the 5% CIH+RH group were dramatically reduced (P<0.01 ). 2) Compared with the control group, the fractalkine expression in the 5%CIH and 5% CIH+RH groups was increased (bothP<0.01). The fractalkine expression in the 5% CIH+RH group was dramatically downregulated compared with that in the 5% CIH group (P<0.01).
Conclusion: CIH can induce liver injury and high fractalkine expression in rat liver tissues.

α-Galactosidase A (α-Gal A ) activities in plasma and peripheral blood granulocytes of 100healthy subjects and one patient with Fabry disease was determined by means of fluorogenic substrate.The results showed that the enzymatic activities of peripheral blood granulocytes and plasma in 100 subjects were (51.97 ± 15.24)and(148.08±26.30) nmol · h-1 · ml-1 respectively.The α-Gal A activities in plasma and granulocytes were positively correlated( r=0.533,P＜0.01 ).The enzymatic activities in peripheral blood granulocytes and plasma of the patients with Fabry disease were 1.05 and 10.06 nmol · h-1 · ml-1 respectively,both much lower than those of 100healthy subjects.These results suggest that α-Gal A activity in plasma and peripheral blood granulocytes can be used for diagnosis and screening of Fabry disease.

Objective To analyze the association between mtDNA mutations and photodamagc after ultraviolet B (UVB) irradiation. Methods Primary human skin fibroblasts (HSF) and primary human epi- dermal keratinocytes of adult (HEKa) were irradiated by sub-lethal doses of UVB thrice a day for 4-5 days. Thereafter, genomic DNA was extracted from irradiated cells and conventional PCR was applied to detect the frequency rates of 4977 bp and 3895 bp mtDNA deletion. To quantitatively analyze the mutation levels, SYBR Green real-time PCR method was performed. Results In both cell lines, the frequency rates and relative copy number of deletions increased with the cumulative doses of UVB exposure (P<0.05). The prevalence rate of 3895 bp deletion peaked 53.3% and and relative copy number reached (49.63±4.38)×10-5, showing a more intense response to the accumulation of UVB radiation than 4977 bp deletion. In HSF, the minimum cumu- lative dose of UVB radiation was 150 mJ/cm2 for the induction of 3895 bp deletion, and 200 mJ/cm2 for the induction of 4977 bp deletion. It seemed that mtDNA deletion was more readily to be induced by UVB radia- tion in HSF than in HEKa. Conclusions The development and accumulation of mtDNA mutation are intimately related with cumulated UVB dose received by skin cells, and the 3895 bp deletion is more reliable in moni- toring the photodamage caused by UV than 4977 bp deletion. Therefore, the 3895 bp deletion may serve as a biomarker for the detection of photodamagc in skin cells. HSF appear to have an increased susceptibility to UVB radiation, which results in a higher frequency and level of mtDNA mutations compared with HEKa.

Thyroid function ultimately depends on appropriate iodine supply to the gland. Thyroid hormone deiodination is an intrinsic component of the thyroid hormone homeostasis. Type Ⅰ iodothyronine deiodinase (D1) plays an important role in thyroid hormone metabolism and has close relationship with thyroid function. Based on successfully establishing animal models of iodine deficiency and iodine excess in Babl/c mice (Babl/c mice were randomly divided into five groups: low iodine (LI), normal iodine (NI), five-fold iodine (5HI) , ten-fold iodine (10HI) and fifty-fold iodine (50HI) group. Three months and six months after admistration, they were sacrificed and thyroids were excised), the expression level of D1 mRNA were examined by using real time quantitative PCR method. D1 activity was analyzed by 125I-rT3 as substrate combined with ion-exchange chromatography. The thyroid hormone was measured with radioimmunoassay method. The data revealed that in the case of iodine deficiency, both D1 mRNA expression and D1 activity was greatly increased(compared with NI groups, P