Objective To explore the effects of gossypol acetate on the morphological features and the gene expression in the femoral head of Sprague-Dawley rat in vivo after treated with dexamethasone .Methods Dexamethasone(Dex) was injected into the abdominal cavity of SD rats at an dose of 10 mg/kg ,twice a week ,and feed gossypol acetate 5 mg · kg -1 · d-1 .The controls re-ceived saline 2 mL injection .The treatment lasted for 12 and 20 weeks .The slices of the femoral head were made for HE and immu-nohistochemical study .The total mRNA was extracted for RT-PCR assessment .Results The cancellous bone trabecular became sparse ,trabecular bone area ratio decreased ,bone marrow fat tissue increased .These changes were fitted for pathological character of bone necrosis .The gossypol acetate could not affect the pathological changes .The proportion of the positive stained osteoblasts increased ,adipocytes decreased .PPARγ,C/EBPα,11β-HSD1 expression enhanced ,Runx2 down regulated in the treatment groups and GAA group .Conclusion Dex can induce evident pathological changes conform to the characters of femoral head necrosis .They may have closed correlation between 11β-HSD1 and the gene expression .But GAA could not affected the pathological changes and abnormality of the gene expression .

Neural stem cells were labeled with superparamagnetic iron oxide (SPIO) and tracked by MRI in vitro and in vivo after implantation. Rat neural stem cells were labeled with SPIO combined with PLL by the means of receptor-mediated endocytosis. Prussian blue staining and electron microscopy were conducted to identify the iron particles in these neural stem cells. SPIO-labeled cells were tracked by 4.7T MRI in vivo and in vitro after implantation. The subjects were divided into 5 groups, including 5 x 10(5) labeled cells cultured for one day after labeling, 5 x 10(5) same phase unlabeled cells, cell culture medium with 25 mug Fe/mL SPIO, cell culture medium without SPIO and distilled water. MRI scanning sequences included T(1)WI, T(2)WI and T(2)*WI. R(2) and R(2)* of labeled cells were calculated. The results showed: (1) Neural stem cells could be labeled with SPIO and labeling efficiency was 100%. Prussian blue staining showed numerous blue-stained iron particles in the cytoplasm; (2) The average percentage change of signal intensity of labeled cells on T(1)WI in 4.7T MRI was 24.06%, T2WI 50.66% and T(2)*WI 53.70% respectively; (3) T2 of labeled cells and unlabeled cells in 4.7T MRI was 516 ms and 77 ms respectively, R(2) was 1.94 s(-1) and 12.98 s(-1) respectively, and T(2)* was 109 ms and 22.9 ms, R(2)* was 9.17 s(-1) and 43.67 s(-1) respectively; (4) Remarkable low signal area on T(2)WI and T(2)*WI could exist for nearly 7 weeks and then disappeared gradually in the left brain transplanted with labeled cells, however no signal change in the right brain implanted with unlabeled cells. It was concluded that neural stem cells could be labeled effectively with SPIO. R2 and R(2)* of labeled cells were increased obviously. MRI can be used to track labeled cells in vitro and in vivo.

Objective To evaluate effects of camptothecin on the autophagy of HaCaT cells.Methods Some cultured HaCaT cells were divided into several groups to be treated with camptothecin at concentrations of 5,10,25,50,100 and 200 nmol/L,and 0.1％ dimethyl sulfoxide (DMSO) (control group),respectively.Cell counting kit-8 (CCK-8) assay was conducted to estimate the proliferative activity of HaCaT cells after 24-and 48-hour treatment,flow cytometry to evaluate cell apoptosis after 24-hour treatment,and Western blot analysis to measure the expression of autophagy-related proteins microtubuleassociated protein 1 light chain 3 (LC3) and p62.Some HaCaT cells were divided into 2 groups to be treated with 10 nmol/L camptothecin and 0.1％ DMSO for 24 hours,respectively.Then,indirect immunofluorescence assay (IFA) was performed to determine the LC3 expression.Results Camptothecin at low concentrations of 5 and 10 nmol/L had no significant effects on the proliferation and apoptosis of HaCaT cells.Compared with the control group,the cellular proliferative rates were significantly inhibited by (31.23 ± 1.00)％,(54.21 ± 8.10)％ and (66.75 ± 10.70)％ in the 50-,100-and 200-nmol/L camptothecin groups after 24-hour treatment respectively,and by (25.81 ± 5.99)％,(44.35 ± 5.32)％,(65.81 ± 8.28)％ and (73.23 ± 9.59)％ in the 25-,50-,100-and 200-nmol/L camptothecin groups after 48-hour treatment respectively (all P ＜ 0.001).After 24-hour treatment,the apoptosis rates were significantly higher in the 50-,100-and 200-nnol/L camptothecin groups (14.46％ ± 2.38％,19.15％ ± 1.59％,29.88％ ± 1.37％,respectively) than in the control group (3.80％ ± 0.13％,all P ＜ 0.001).After 24-hour treatment with 5 and 10 nmol/L camptothecin,the protein expression of LC3 Ⅱ was significantly up-regulated,while p62 protein expression was significantly down-regulated:IFA showed that the percentage of autophagosome-positive cells was significantly higher in the 10-nmol/L camptothecin group than in the control group after 24-hour treatment (36.67％ ± 4.55％ vs.6.23％ ± 0.92％,t =6.546,P =0.003).Conclusions Camptothecin at low concentrations of 5 and 10 nmol/L can induce autophagy of HaCaT cells,but has no obvious effects on cell proliferation and apoptosis.Camptothecin at concentrations of 50,100 and 200 nmol/L can inhibit cell proliferation,promote cell apoptosis,and decrease autophagy levels.

OBJECTIVETo investigate the effect of chronic endurance exercise on microcirculatory reserve capacity of biceps brachii in Chinese rowers and provide a certain basis for the date standard foundation of monitoring of functional status and the foundation of database of reserve capacity of blood of Chinese rowers.

METHODSEmpty stomach in the morning, 77 rowers from different groups and 24 common health people were noninvasive tested by using PeriFlux System 5000, the test indexes include the microcirculatory reserve capacity and other related indexes of biceps brachii. The test sites of all athletes were the same space in biceps brachii of the right side of body, there was no space differences of all athletes . All athletes were tested in the relatively stable functional status, common people were healthy. The test value included basic values and heating values, put the before and after heating of microcirculatory blood perfusion (MBP) as the microcirculatory reserve capacity.

RESULTSHeavyweight female (198. 97 ± 98. 81) > heavyweight male (183. 45 ± 64. 31) > lightweight male (151. 01 ± 65. 96) > lightweight female(140.53 ± 43.22) > common male people(127.21 ± 56.38) > common female people(103.54 ± 33.41), the microcirculatory reserve capacity of each group athletes were higher than common people, except the comparison between lightweight female and common male people, and there was no significant difference among the different group athletes.

CONCLUSIONChronic endurance exercise can improve the microcirculatory reserve capacity of rowers, especially the heavyweight rowers; the normal value of microcirculatory reserve capacity of heavy weight rowers should be more than 160, and lightweight rowers should be more than 120. There was no significant difference among different sex athletes, if the value of microcirculatory reserve capacity is significant lower than normal, it shows that athletes are in the state of fatigue.

Arm ; Athletes ; Female ; Humans ; Male ; Microcirculation ; Muscle, Skeletal ; blood supply ; Sports

Objective To investigate altered microstructural integrity in acute carbon monoxide(CO)intoxication by diffusion tensor imaging (DTI).Methods A total of 25 CO intoxication patients (5.0±1.44 post-intoxication)and 37 healthy volunteers matched with age, sex and educational level were involved and underwent DTI.The fractional anisotropy (FA)and apparent diffusion confficient (ADC) of total 26 regions of bilateral cerebellum,substantia nigra,hippocampus,white matter of frontal lobe,head of caudate nucleus,pallidum,thalamus, anterior limb of internal capsule,posterior limb of internal capsule,white matter of occipital lobe,white matter of parietal lobe,and genu of corpus callosum,splenium of corpus callosum were measured.Compared the patients against control for FA and ADC by paired t-test.Results FA of patients with CO intoxication decreased in bilateral pallidum,anterior limb of internal capsule,substantia nigra,right cerebellum,left below frontal white matter,right frontal white matter and genu of corpus callosum (P <0.05).ADC decreased in right substantia nigra,and left pallidum (P <0.05).ADC increased in right frontal white matter and bilateral occipital lobe white matter (P<0.05).Conclusion There is loss of microstructural integrity in acute CO intoxication,perhaps representing the underlying mechanism of delayed encephalopathy after carbon monoxide poisoning.

Objective To investigate the efficacy and side effects of the molecular targeted drug sunitinib in the treatment of metastatic renal cell carcinoma(mRCC). Methods Fifteen patients with histopathological confirmed mRCC,including 10 males and 5 females,were enrolled in the study.The median age was 56(range from 37 to 73 years).Fourteen cases of clear-cell RCC and 1 papillary RCC were diagnosed.Thirteen patients had prior nephrectomy,and 6 patients were treated with cytokines previously.All of the patients were given sunitinib at a dosage of either 50 mg daily(4 weeks on/2 weeks off)or 37.5 mg daily continuously.Objective response rate(ORR),progressive-free survival(PFS)and overall survival(OS)were evaluated,and adverse events were also observed. Results The median follow-up was 13 months(2-24 months).According to RECIST,partial response could be evaluated in 8 patients and stable was found in another 5 patients.Only 2 patients were confirmed disease progression.The ORR was 53% with the disease control rate of 87%,However,the median PFS and OS were not yet available due to the short-term follow-up.During the treatment,the most common adverse events experienced by patients were hand-foot syndrome 11/15(73%),changes in hair color 10/15(67%),mucositis 9/15(60%),alopecie 9/15(60%),diarrhea 8/15(53%)and neutropenia 8/15 (53%). All of adverse events were manageable and reversible. Conclusions Sunitinib as a single agent in the treatment for Chinese mRCC patient is efficient and the side effects are reversible. Further long-term follow-up and expanded samples should be expected to confirm the efficacy and safety of sunitinib.

AIM:To investigate the expression of transmembrane protein 16A(TMEM16A) in Fischer rat thy-roid follicular epithelial ( FRT) cells and its electrophysiologic properties .METHODS: The eukaryotic expression vector of pUB6/V5-TMEM16A was constructed and transfected into FRT cells by liposome-mediated transfection .In order to ob-tain the high efficiency of gene transfection and expression , the quantity and ratio of lipid/DNA complexes were optimized . The FRT cells stably expressing TMEM16A were gained by the selection with blasticidin and confirmed by the techniques of RT-PCR and immunofluorescence .The expression and location of TMEM 16A in the FRT cells were observed under an in-verted fluorescence microscope .TMEM16A protein was associated with calcium-dependent chloride current , as measured with halide-sensitive fluorescent protein and patch-clamp technique .RESULTS: The results of double digestion and se-quencing indicated that TMEM16A was cloned into pUB6/V5.The results of RT-PCR and immunofluorescence confirmed that TMEM16A was expressed in the FRT cells after transfection with TMEM16A.The classical calcium-activated chloride channel currents were recorded in the FRT cells stably expressing TMEM 16A by the technique of patch-clamp and halide-sensitive fluorescent protein YFP-H148Q/I152L.CONCLUSION:The protein expression of TMEM16A in the FRT cells was observed.TMEM16A is the molecular identity of calcium-activated chloride channels .

OBJECTIVETo analysis the outcomes and the technique of surgical operation of osteoporotic ankle fracture in the elderly.

METHODSOne hundred and twenty-eight patients with osteoporotic ankle fracture were treated by surgical operation from January 2003 to December 2008, included 26 males and 102 females, with an average age of 69 years old ranging from 50 to 82 years. According to Lange-Hanson classification, there were 113 cases of supinatio-extorsion, 18 of pronate-extorsion, 5 of supinatio-adduction, 2 of pronate-abduction. The methods of fixation, the complications, and the AOFAS scores after operation were recorded.

RESULTSAll the 128 patients were followed up from 6 months to 6 years (averaged in 36 months). Delayed wound healing occurred in 11 cases (8.6%). There was no deep infection, nonunion of the fracture, and revision. The average AOFAS scores after operation was 83.6 +/- 6.3 (the mean pain scores were 33.5 +/- 5.6, the mean function scores were 40.2 +/- 7.4, the mean alignment scores were 10).

CONCLUSIONSurgical treatment should be the first choice in osteoporotic ankle fracture in most cases. Improved procedures can make the results better.

Aged ; Aged, 80 and over ; Ankle Injuries ; surgery ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Osteoporotic Fractures ; surgery ; Retrospective Studies

BACKGROUND: Hydroxyapatite ceramic is one of the bioactive ceramics commonly used in clinic. Accurate regulation of the surface microstructure of hydroxyapatite ceramics is favorable for improving its biological performance, which is needed to be investigated thoroughly. OBJECTIVE: To investigate the effect of surface roughness on the wettability and protein adsorption ability of hydroxyapatite ceramic as well as mesenchymal stem cell proliferation on its surface. METHODS: Hydroxyapatite powders were synthesized by chemical precipitation method. Hydroxyapatite ceramics were produced by cold isostatic pressing of the powders and then high-temperature sintering of the obtained green compacts. After the different grinding and polishing processes, the hydroxyapatite ceramics with three distinct surface roughness were fabricated. The phase composition, microstructure, surface roughness and water contact angle of the hydroxyapatite ceramics were evaluated. Bovine serum albumin was used as the protein model to study the effect of surface roughness on protein adsorption. The in vitro cell culture was carried out to investigate the effect of surface roughness of hydroxyapatite ceramics on the growth and proliferation of rat bone marrow mesenchymal stem cells. RESULTS AND CONCLUSION: The hydroxyapatite ceramics with no surface treatment had higher surface roughness, presenting with marked undulating surface; the hydroxyapatite ceramics with surface grinding had a reduction in the surface roughness, but still had an undulating surface; and the hydroxyapatite ceramics with polishing treatment had a smooth surface. The surface roughness of the three hydroxyapatite ceramics with different surface treatment as mentioned above was 448.4, 229.9 and 18.6 nm, respectively, and there was a significant difference between them. The results of contact angle tests indicated that water contact angle of the hydroxyapatite ceramic was influenced by its surface roughness. Lower surface roughness meant larger contact angle, leading to the decreased hydrophilicity of the hydroxyapatite ceramics. The protein adsorption experiment showed that the hydroxyapatite ceramics were favorable for bovine serum albumin adsorption, and the amount of the adsorbed bovine serum albumin was dependent on the surface roughness of the ceramic as well as the initial concentration of the protein solution. The hydroxyapatite ceramics with higher surface roughness would absorb more bovine serum albumin, and the bovine serum albumin adsorption increased with the increasing initial concentration of the protein solution. The in vitro cell culture confirmed that surface roughness impacted the proliferation of rat bone marrow mesenchymal stem cells. Although the good attachment and growth of bone marrow mesenchymal stem cells were observed on the three hydroxyapatite ceramics, faster proliferation of mesenchymal stem cells occurred on the hydroxyapatite ceramics with lower and nano-scaled surface roughness was observed. Therefore, the surface roughness of hydroxyapatite ceramics exerts great effects on hydrophobicity, protein adsorption, and cell proliferation and growth.

In this study, Streptomyces avermitilis Bjbm0006 which produces four avermectin B components was used as an original test strain. A replacement plasmid containing a gene cluster bkdAB (branched-chain alpha-keto acid dehydrogenase gene) involved in the biosynthesis of avermectin B in S. avermitilis Bjbm0006 was constructed by means of PCR technique and then named as pHJ5821 (pHZ1358::bkdAB&erm). A recombinant strain Bjbm5821 was obtained after the gene cluster was interrupted by double crossover. This strain was tested in laboratory conditions and analysed by PCR using the total DNA as template. The HPLC analysis showed that the strain Bjbm5821 synthesized the same 'a' components Bla and B2a as the original strain did. However, It lost the ability for the production of 'b'components for example B1b and B2b. A novel compound was detected in fermentation products. The results of present study suggests that the production of gene cluster bkdAB may play a main role similar to alpha-ketoisovaleric acid dehydrogenase in the pathway of avermectin synthesis.
3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide) ; genetics ; Ivermectin ; analogs & derivatives ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Streptomyces ; enzymology ; genetics