To study the apoptotic regularity of peripheral lymphocytes in radiation compound wound healing and explore their relationship to radiation delayed wound healing, apoptosis and Bax and Bcl 2 proteins were estimated by in situ terminal labeling (TUNEL) and immunohistochemical method of alkaline phosphatase, respectively. White blood cells in radiation compound wound(RW) group decreased and reached the lowest 3 days after wound, it was still lower in RW group than in simple wound(SW) group 5 days after wound; apoptosis of peripheral lymphocytes of both groups increased, however, the lymphocyte apoptosis in RW group was higher than that in SW from beginning to end in the whole process of wound healing; the expression of Bax protein of lymphocyte increased at different times after wound, and the expression in RW group was obviously higher than that in SW group, while Bcl 2 showed an opposite tendency. It is suggested that reduction of white blood cells and increment of lymphocyte apoptosis are the major causes of radiation delayed wound healing; Bax and Bcl 2 proteins take part in the regulation of lymphocyte apoptosis in wound healing.

To investigate the culture method for epidermal stem cell in vitro. Rat skin was peeled from the subcutaneous tissue gently, and cut into small pieces, then incubated for 24h in 0 25%trypsin in a 4℃ shaker. The epidermis was separated from the dermis, and shaken for 10min in 0 25% trypsin at 37℃ to dissociate into single cells. Digestion was terminated by the addition of culture medium +10% serum, and the cells were gently centrifuged and resuspended in the culture medium, which constituted EMEM without calcium, supplemented with 0 05mmol/L CaCl 2 , 9% chelexed FBS, 4ng/ml EGF, 0 5?g/ml gentamicin, and 50% fibroblast conditioned medium (CM ). The CM ( EMEM without calcium, 0 05mmol/L CaCl 2 , 9% chelexed FBS, 0 5?g/ml gentamicin ) was collected from freshly isolated primary neonate fibroblast cultures after 48h, which were passed through a filter with 0 45?m pores, and stored at -20℃ for use. 1?10 6 /ml epidermal cells were incubated for 10～15min at 37℃ on dishes coated with collagen Ⅳ, then the nonadherent cells were rinsed off. The adherent cells were grown to confluency. Cultures were subcultured in a solution of TGG for 5～10min. Once free of the dish, cells were centrifuged, resuspended in fresh medium, and then cultured at 1?10 5 /ml. Cultures were observed for colony formation under a phase contrast microscope, and the structure of the rapidly adherent cells was observed under electron microscopy; the expression of ? 1 integrin and K19 was detected in the rapidly adherent cells and colony cells with SP immunohistochemical methods. The results showed that the cells selected by rapid adherence to collagen type Ⅳ formed large colonies at 7 days, and showed a im mature feature under electron microscopy, manifesting ? 1 integrin and K19 expression. It is suggested that epidermal stem cells could be cultured in vitro .

The development of sweat glands is a very complex biological process, which involves many factors. In this study, the correlation between epidermal growth factor (EGF), matrix metalloproteinases (MMP 2,MMP 7) and development of sweat glands in human embryos was explored. Furthermore, the mechanism(s) underlying the induction of epidermal stem cells into sweat gland cells was elucidated. Skin biospies of human embryos obtained from spontaneous abortions at different gestational ages from 11 to 31 weeks were used. The dynamical expression of EGF, MMP 2, MMP 7 and keratin 7(K7) in developing sweat gland cells or extracellular stroma surrounding the sweat gland cells were examined with S P immunohistochemical methods. The localization of the cellular sources of MMP 2 and MMP 7 was examined with in situ hybridization. The results showed that at 14 20 wk of gestation, a gradual increase of EGF immunoreactivity was observed not only in developing sweat gland buds but also in extracellular stroma surrounding the buds,and the expression intensity peaked at 20～22 wk of gestational age. All mRNA positive buds or cells in developing sweat glands contained corresponding immunoreactive proteins. The immunostaining for K7 appeared in early sweat gland buds at 14～16wk of gestation, and from then on, K7 was concentrated in developing sweat gland cords or cells. It is suggested that the morphogenesis of sweat gland in human fetal skin begins at 14～16wk of gestational age, and essentially completes by 24wk. There is a close relationship among EGF ,extracellular matrix remodeling and morphogenesis of sweat glands, and EGF is one of the inducers in the development and maturity of sweat gland buds or cells.

Objective:To investigate the protective effect of pine pollen on lipopolysaccharide (LPS)-induced learning and memory impairments in mice and the underlying mechanism.Methods:Sixty mice were randomly divided into four groups ( n = 15/group): normal control, model, low-dose pine pollen (500 mg/kg) and high-dose pine pollen (1 000 mg/kg). Mouse models of learning and memory impairment were established by lateral ventricle injection of LPS. The learning and memory abilities of mice were determined by the Morris water maze test. Superoxide dismutase (SOD) activity and glutathione (GSH) and malondialdehyde (MDA) levels in the hippocampus of mice were measured. Interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), dopamine (DA), and norepinephrine (NE) levels in the hippocampus were also determined. Results:The latency in the passive avoidance test in the model group was significantly shorter than that in the normal control group [(134.80 ± 33.89) s vs. (282.20 ± 17.43) s, t = 4.23, P < 0.01]. The number of errors in the model group was significantly higher than that in the normal control group [(4.00 ± 1.58) vs. (1.20 ± 1.30) times, t = 2.85, P < 0.01]. The latency in the passive avoidance test in the low-dose pine pollen (500 mg/kg) and high-dose pine pollen (1000 mg/kg) groups was significantly longer than that in the normal control group [(189.40 ± 27.21) s or (213.40 ± 21.26) s vs. (134.80 ± 33.89) s, t = 3.21, 4.38, all P < 0.05]. The number of errors in the low-dose pine pollen (500 mg/kg) and high-dose pine pollen (1 000 mg/kg) groups was significantly lower than that in the normal control group [(1.60 ± 1.44) times or (1.40 ± 1.44) times vs. (4.00 ± 1.58) times, t = 5.12, 6.42, both P < 0.05]. SOD activity and GSH, DA and NE levels in the hippocampus in the model group were significantly decreased compared with the normal control group [SOD: (7.59 ± 1.77) kU/g vs. (39.90 ± 6.37) kU/g; GSH: (3.49 ± 0.13) mmol/g vs. (6.37 ± 0.14) mmol/g; DA: (418.42 ± 2.57) ng/L vs. (586.37 ± 3.64) ng/L; NE: (187.20 ± 5.41) ng/L vs. (298.42 ± 2.32) ng/L, t = 3.67, 8.23, 2.23, 3.65, all P < 0.05]. MDA, IL-6 and TNF-α levels in the hippocampus in the normal control group were significantly higher than those in the model group [MDA: (8.79 ± 0.82) mmol/g vs. (2.62 ± 0.16) mmol/g, IL-6: (48.07 ± 5.56) ng/L vs. (18.76 ± 1.42) ng/L, TNF-α: (87.20 ± 4.31) ng/L vs. (22.42 ± 3.39) ng/L, t = 7.45, 2.67, 4.35, P < 0.05 or P < 0.01]. SOD activity, GSH, DA and NE levels in the hippocampus in the low-dose pine pollen (500 mg/kg) and high-dose pine pollen (1 000 mg/kg) groups were significantly higher than those in the model group [SOD: (18.80 ± 2.39) kU/g, (28.70 ± 2.36) kU/g vs. (7.59 ± 1.77) kU/g, GSH: (5.04 ± 0.36) mmol/g, (5.45 ± 0.17) mmol/g vs. (3.49 ± 0.13) mmol/g, DA: (488.37 ± 3.46) ng/L, (506.29 ± 5.72) ng/L vs. (418.42 ± 2.57) ng/L, NE: (225.65 ± 3.72) ng/L, (239.76 ± 5.58) ng/L vs. (187.20 ± 5.41) ng/L, t = 4.56 or 6.71, t = 4.65 or 5.32, t = 4.73 or 6.72, t = 3.84 or 5.63, P < 0.05 or P < 0.01]. MDA, IL-6 and TNF-α levels in the hippocampus in the low-dose pine pollen (500 mg/kg) and high-dose pine pollen (1 000 mg/kg) groups were significantly lower than those in the model group [MDA: (5.72 ± 0.47) mmol/g, (3.77 ± 0.23) mmol /g vs. (8.79 ± 0.82) mmol/g, IL-6: (28.42 ± 3.54) ng/L, (23.43 ± 5.62) ng/L vs. (48.07 ± 5.56) ng/L, TNF-α: (48.87 ± 4.82) ng/L, (39.65 ± 6.69) ng/L vs. (87.20 ± 4.31) ng/L, t = 6.31 or 7.28, t = 3.46 or 6.31, t = 4.28 or 3.57, P < 0.05 or P < 0.01]. Conclusion:Pine pollen can improve LPS-induced learning and memory impairments possibly through up-regulating the levels of monoamine neurotransmitters DA and NE and inhibiting the levels of oxidative stress and inflammatory reaction in the hippocampus of mice.

ObjectiveTo explore the morbidity rate of deviation of nasal septum about personnels who take part in physical examination in the enterprise and facilities of Qinhuangdao,and clinic symptom.MethodsTransverse questionnaire investigation and normal physical examination were adopted.2604 personnels who take part in physical examination were choiced.ResultsMorbidity rate of deviation of nasal septum was 17.6％.The rate of men' s exceeds that of women' s.ConclusionMorbidity rate of deviation of nasal septum was rather high,clinic symptom of snuffle was primary.

Objective To compare the efficacy of self-made bone cement spacers containing vancomycin(vancomycin spacer)and Link infection-treating spacers(Link type spacer)for chronic infection after total hip arthroplasty(HTA)and investigate the choice of spacers for the two-stage revision of infection.Methods The study included 26 patients with infection combined with HTA,aged 32-82 years(mean 69.5 years).In the first stage,after complete debridement and removal of infected prostheses,the vancomycin spacers were implanted in 14 patients,while the Link type spacers were implanted in the rest 12 patients.Two weeks of intravenous administration of antibiotics and then six weeks of oral administration of antibiotics were done postoperatively.White blood cells(WBCs),erythrocyte sedimentation rate(ESR)and C-reactive protein(CRP)were measured periodically after operation.THA revision was performed in the second stage.Clinical outcomes were evaluated according to the pre-and post-operative Harris hip score.Results All patients obtained primary healing of incision after complete debridement,removal of prostheses and implant of the vancomycin spacers or the Link type spacers.Two-stage revision surgery was performed at 3-11.5 months after the one-stage surgery(mean 5.2 months).The follow-up ranging from 12 to 33 months(mean 20.8 months)manifested no signs of reinfection.The mean Harris hip score was increased from 27.9 points before one-stage operation to 52.9 points before two-stage operation,and 83.6 points at the latest follow-up(P＜0.05)One of 12 vancomycin spacers was broken before the revision surgery,as was free from the Link spacers.Conclusions(Ⅰ)Two-stage revision is an effective method in the treatment of infection after HTA because of high eradication rate of infection,effective pain relief and good postoperative functional result.(2)The vancomycin spacers and the Link type spacers can both effectively control infection and restore part of limb function.(3)The Link type spacers take advantages of simple in operation and the function improvement of the limbs during the intermittent period of treatment,but disadvantages of the catheters being prone to block,limited slow release of antibiotics and high cost of treatment.Thereby,the Link type spacer is suitable for the patients with relatively strong constitution,little serious infection,or high requirement of limb functions at the interval of two operations.

Objective To explore the treatment using two different ventilation tubes insertion for secretory o-titis media ,and to determine which factors are associated with the tympanic membrane perforation after a extrusion or surgical removal of a ventilation tube(VT) .Methods A retrospective analysis of 121 cases (210 ears) with re-fractory secretory otitis media from 1998 June to 2013 March was carried out .There were 58 cases (105 ears ) and 63 cases (105 ears) in patients and control group ,respectively .All subjects had more than 2 years of history ,and were treated by two types of ventilation tube (Goode -T and Shepard tube) .Various factors were analyzed to deter-mine the factors associated with persistence of a tympanic membrane perforation after VT extrusion or removal ,in-cluding gender ,age ,medical history ,ventilation tube pattern ,location ,history of previous VT insertions ,intuba-tion period reason for VT insertion ,condition of the TM ,nature of the tympanic cavity effusion and previous ade-noidectomy ,et .Results Analysis factors of perforation group (105 ears) and the control group (without perfora-tion ,105 ears) ,were statistically significant in ventilation tube pattern (χ2 =6 .916 ,P<0 .01) ,history of previous VT insertions(χ2 =7 .325 ,P<0 .01) ,frequency of previous VT insertions (χ2 =13 .01 ,P<0 .01)and the nature of the tympanic cavity effusion(χ2 =7 .04 ,P<0 .01) .The proportion of perforation group about Goode - T ventilation tube ,history of previous VT insertions ,the frequency of tube (more than 3 times) ,mucus of the middle ear cavity were 58 .4% ,57 .3% ,66 .7% and 57 .9% respectively ,others had no significant difference between the two groups . The multivariate analysis showed that ,regardless of the ventilation tube ,the frequency of tube was the tympanic membrane perforation risk factor ;Goode-T tube increased the risk of perforation of tympanic membrane was 1 .637 times than Shepard tube(OR=1 .637 ,P<0 .01) .Conclusion Using the Goode - T type ventilation tube ,history of previous VT insertions ,the frequency of tube (more than 3 times) ,mucus of the middle ear cavity were four risk factors for persistence of a tympanic membrane perforation after VT insertion for secretory otitis media .

OBJECTIVE: To evaluate the effect of integrated traditional Chinese and Western medicine therapy on the progression of acute renal failure in patients with chronic renal insufficiency. METHODS: Thirty-two patients with chronic renal insufficiency developed acute renal failure recently were treated with Chinese herbs and western drugs intravenously and clysterizing of Chinese herbs liquid for 30 minutes, and the treatment course was 14 days. Assessment of liver and renal function, blood routine, electrolytes and endogenous creatinine clearance rate (Ccr) was performed before and 2 weeks after the treatment. RESULTS: The levels of hemoglobin (HB), white blood cell count (WBC) and serum electrolytes showed no significant changes after the treatment. The levels of blood urea nitrogen (BUN) and serum creatinine (SCr) decreased, while the level of Ccr increased significantly (P<0.05) after the treatment. The total effective rate was 65.6%. CONCLUSION: The integrated traditional Chinese and Western medicine therapy can effectively delay the deterioration of renal function in patients with chronic renal insufficiency accompanied by acute renal failure.

Objective To investigate the effect of parathyroid hormone (PTH) on the transition and connective tissue growth factor (CTGF) expression of human renal proximal tubular epithelial cell line HK-2 . Methods The expression of CTGF mRNA and protein of HK-2 cells were measured by real time RT-PCR and Western blot respectively . The effect of PTH on the phenotypic transformation of HK-2 cells was examined by light microscopy . The expression of α-smooth muscle actin (α-SMA) in HK-2 cells was detected by immunofluorescence . Results Basal level of CTGF mRNA and the protein expression were detected in HK-2 ceils . PTH upregulated the expression of CTGF mRNA and protein with the maximal response at the concentration of 10-10 mol/L and the best stimulating time was at 72 h . After exposure to PTH (10-10tool/L) for 12 hours, the highest level of luciferase activity was 1 .96 fold as compared to control (1 .888±0 .078 vs 0 .989±0 .030, P＜0 .01 ) . Untreated cells showed negligible expression of ±-SMA,whereas ±-SMA expression was significantly increased in cells treated with PTH . Conclusion PTH up-regulates CTGF expression and induces transition of HK-2 cells .

Using the cyclodextrin bonded polysiloxane (bikis〔（2,6-di-O-pentyl-3-O-hex-6-O-enyl)-pentakis(2,6-di-O-pentyl-3-O-methyl)-β-cyclodextrm-polysiloxane〕) as gas chromatographic stationary phase, α-hydroxy acetones were separated and the values of the enantiomeric excess(e.e) of 3-hydroxy-2-butanone were determined. The results showed that the high resolution gas chromatography (HRGC) using the chiral gas chromatographic stationary phase could determine the productive rate of the asymmetric hydrogenaton reaction and evaluate the enantoselectivity of the catalyst system.