Objective To approach the design and clinical application of large anterolateral thigh flap and its effect in wound repair.Methods The flaps were designed according to the anatomical features of perforating branches in the anterolateral thigh flaps.When a flap was chipped,a thick branch or a terminal branch of original vessel was reserved,another suitable perforating branch was selected in the proximal or distal end of the flap,and then the two vessels were anastomosed to enlarge the range of blood supply.If the vessel pedicle of a flap was a musculocutaneous perforating branch,the perforating branch of anastomosis was cut at out-point of muscle.If the vessel pedicle of a flap was a intermusclar branch or a direct skin artery,the perforating branch of anastomosis was cut widely.From May 2006 to May 2012,the technique was applied in 28 patients with large skin defect of limbs.The diameters of perforating branches obtained at out-point of muscles were measured during surgery.The survival of flaps was observed after surgery and complications in donor sites were checked during follow-ups.Results There were 18 flaps whose vessel pedicle were musculocutaneous perforating branches.The branches were cut at outpoint of muscles.The diameters of these vessels were measured during surgery.They ranged from 1.3 mm to 1.8 mm with an average of 1.45 mm.All of the vessels could be anastomosed.All 28 flaps survived.All flaps survived.The areas of the flaps ranged from 22 cm × 15 cm to 42 cm × 14 cm.Artery crisis happened in 2 flaps whose vessel pedicle were musculocutaneous perforating branches.The second look operation found that the areas of artery anastomosis of perforating branches and vessel pedicles were compressed by hematoma and thrombus formed.The 2 flaps survived after the hematoma was cleared away and the vessels were reanastomosed.There were no infections.Both the donor and recipient site healed by first intention with no necrosis of flap margin.All 28 patients were followed up by 4-13 months with an average of 8 months.There were no apparent collapse deformities,muscle necrosis,declines of muscle strength and muscle hernia in the donor sites.The appearance of flaps was flat,the color was close to normal and the quality was fine.Conclusion It is a safe and effective method to repair wound surface by large anaterolateral thigh flap obtained by the modus operandi of perforating branch anastomosis.

OBJECTIVE: To investigate the effects of arc-track private lock pedicle orthopedic fixation system Ⅱ (ALPF Ⅱ) on treatment of spinal diseases. METHODS: A total of 86 patients with spinal diseases were treated using self-made ALPF in the Wendeng Orthopaedic Hospital and were included in this study. Of these patients, 14 suffered from cervical spinal stenosis complicated by cervical vertebral destabilization, 29 from thoracolumbar fractures and dislocations, 15 from lumbar spinal stenosis complicated by lumbar vertebral destabilization, 2 from lumbar spondylolisthesis, 8 from spinal tuberculosis, 6 from ankylosing spondylitis, 9 from idiopathic scoliosis, and 3 from congenital scoliosis. According to conditions, different therapeutic regimens were selected. Postoperatively, regular follow-up was performed to observe vertebral healing, intervertebral height, spinal column sequence, and neurological function recovery. RESULTS: All patients were followed up for 9-30 months (average 12 months). The improvement rate of neurological function, spinal mobility, back pain, and melosalgia was 94.1%, 65.9%, 92.1%, and 87.4%, respectively. The postoperative anterior and posterior vertebral heights were apparently recovered, and kyphotic angle was well corrected. No screw, rod loosening or breakage was found. CONCLUSION: Self-made ALPF Ⅱ is an internal fixation method for treatment of spinal diseases. It provides good reduction, reliable curative effects, less complications, and no biocompatibilities.

Objective To study the effect of rhu-IFN-? on immune state and vertical transmission in pregnant mice infected with Toxoplasma gondii. Methods Sixty pregnant BALB/c mice were randomly divided into three groups: a control group,infected group and treatment group. In the infected group and treatment group,each mouse was injected with T.gondii tachyzoites peritoneally on the day 8 of gestation. In the treatment group,each mouse was treated with 1 000 U rhu-IFN-? on the day 7,8,9 of gestation. Blood was collected from the tail veins of all the mice on the day 10,12 of gestation. The levels of blood CD4+ and CD8+ T cells were detected by flow cytometry. Meanwhile,on the day 12 of gestation,all mice were anatomized to observe live embryo rate and the infection status in fetal brain tissue. Results Compared with the control group,the levels of CD4+ T cells in the infected group and treatment group were low,and the levels of CD8+ T cells high on the day 10,12 of gestation,so the ratio of CD4+/CD8+ was inversed. However,compared with the infected group,the levels of CD4+ T cells in the treatment group were high and the levels of CD8+ T cells low on the day 10,12 of gestation,so the ratios of CD4+/CD8+ were high on the day 10,12 of gestation. Meanwhile,the live embryo rate was high and the infection rate of intrauterine embryonic low. Conclusion A proper dose of rhu-IFN-? could improve the function of immunity system and reduce the vertical transmission probability in pregnant mice infected with T.gondii.

Objective To explore the function of 5-Aza-CdR in B-cell acute lymphocytic leukemia cell line NALM-6 and its influence on the expression of microRNA (miRNA) in the cells. Methods NALM-6 was treated with different concentrations of 5-Aza-CdR. Cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) test, and DNA methyltransferase (DNMT) mRNA expression level was detected by reverse transcription PCR (RT-PCR). The expression changes of miRNA were detected by miScript miRNA PCR Array chip in cells after methylation. Results NALM-6 cell growth was inhibited by different concentrations of 5-Aza-CdR processing time, reaching to the maximum inhibitory rate was (74.163 ±0.381) %. 5-Aza-CdR affected concentrations was inversely proportional with expression level of DNMT mRNA. After 1 000 μmol/L of 5-Aza-CdR was dealed with cell 72 h, the relative expression of DNMT-1 was reduced to 0.453 ±0.021, DNMT-3L was 0.003±0.001, DNMT-3B was 0.395±0.019. MiScript miRNA PCR array sieved out 3 miRNA (miR-184, miR-23a-3p, miR-34a-5p) associated with DNA methylation. Conclusions 5-Aza-CdR down regulates the expression of DNMT gene in NALM-6 cells, and inhibits the proliferation of cells. MiR-184, miR-23a-3p and miR-34a-5p are related to DNA methylation in the occurrence and development of B-cell acute lymphocytic leukemia.

Objective To study the effect of pioglitazone on the differentiation and function of rat osteoclast-like cells (OLC), and to probe the relationship between activated PPARγ2 and osteoclasts. Methods On day 1 of OLC formation from nonadherent bone marrow ceils (BMC) obtained from rats induced by M-CSF and receptor activator of NF-кB ligand (RANKL), 1, 5 and 10μmol/L pioglitazone hydrochloride was added. RT- PCR was performed to determine the mRNA expressions of PPARγ2 and receptor activator of NF-кB (RANK) on day 3, 5 and 7 during incubation, the number of tartrate-resistant acid phosphatase (TRAP)-positive cells,the number of bone resorption pits and the ratio of its area on dentin slice were counted, the activity of TRAP and the mean fluorescence intensity of integrin β3 (CD61) of OLC were also measured. Results (1) The effect on the differentiation of OLC: The addition of pioglitazone at the start of the culture period induced a dose-dependent decrease in TRAP-positive OLC and the activity of TRAP (P < 0.01 or P < 0.05) ; the mRNA expression of PPARγ2 was up-regulated by 5 and 10 μmol/L pioglitazone in the early stage of incubation and attenuated with thematuration of OLC on the contrary, however, the expression of RANK was down-regulated by 5 and 10 μmol/L piolitazone in every stage of incubation (P < 0.05 or P < 0.01), combined with decrease in TRAP-positive OLC from day 3 by 10 μmol/L pioglitazone. (2) The effect on the function of OLC: the number of bone resorption pits and the ratio of its area on dentin slice were decreased in groups of 5 and 10 μmol/L pioglitazone (P < 0.01 orP < 0.05), no obvious change was noted in the group with 1 μmol/L pioglitazone compared with the control group; the mean fluorescence intensity of CD61 were down-regulated in groups of 5 and 10 μmol/L pioglitazone (P < 0.05 or P <0.01). Conclusion Activation of PPARγ2 pathway by pioglitazone could partially inhibit differentiation and function of OLC derived from rat BMC.

OBJECTIVE To investigate the effect and mechanism of acute exposure to sodium cyanide(NaCN)on brain nerve damage induced by closed hypoxia in mice.METHODS ① Mice were randomly divided into hypoxia+NaCN 0(hypoxia control group),2.56,3.8,and 5.1 mg·kg-1 groups.After ip adminis-tration of different concentrations of NaCN,the mice were immediately placed into a closed hypoxic tank and the hypoxia survival time was observed.②Mice were divided into normal control,NaCN 3.8 mg·kg-1,hypoxia(30 and 60 min)and NaCN 3.8 mg·kg-1+hypoxia(30 and 60 min)groups.After grouping,the pH,oxygen saturation(sO2),oxygen tension(pO2)and carbon dioxide partial pressure(pCO2)of arterial blood of mice were detected using an arterial blood gas analyzer.The cortical cerebral blood flow of mice was detected using a laser speckle imager.The dry and wet brain tissue were weighed separately,and the brain moisture content was calculated.The kit was used to detect the activity of total superoxide dismutase(T-SOD)and the content of malondialdehyde(MDA)in the hippocampus.TUNEL staining was used to detect the apoptosis rate of cells in the hippocampus.HE staining was used to detect path-ological changes in the hippocampus.RESULTS ①Compared with the hypoxic control group,the sur-vival time of mice in the hypoxic+NaCN groups was significantly prolonged(P<0.01).②Compared with the normal control group,the hypoxia 30 min group showed upregulation of arterial blood p CO2(P<0.05),downregulation of p O2(P<0.05).The hypoxia 60 min group showed upregulation of arterial blood p CO2(P<0.05)and downregulation of cortical cerebral blood flow(P<0.05).In the NaCN 3.8 mg·kg-1 group,arterial blood p O2 and s O2 were significantly downregulated(P<0.05),so was cortical cerebral blood flow(P<0.01),but MDA content and T-SOD activity were significantly upregulated(P<0.01),and the brain moisture content was increased(P<0.01).Compared with the hypoxia 30 min group,s O2 and p O2 of arterial blood in the NaCN+hypoxia 30 min group were significantly upregulated(P<0.05),while p CO2 was significantly downregulated(P<0.05).Compared with the hypoxia group at corresponding time points,the NaCN+hypoxia 30 or 60 min groups showed significant downregulation of cerebral blood flow(P<0.01),significant upregulation of MDA content and T-SOD activity(P<0.01),and signifi-cant upregulation of brain moisture content(P<0.01).HE staining results showed that the NaCN 3.8 mg·kg-1 group and the NaCN+hypoxia group(30 or 60 min)showed significant cell swelling and vacuolization in cells in the hippocampal tissue,a decrease in the number of neurons,nuclear pyknosis and deep staining.TUNEL fluorescence results showed that the NaCN 3.8 mg·kg-1 group significantly increased the apop-tosis rate of the mouse hippocampus compared with the normal control group(P<0.05).The NaCN+ hypoxia 30 and 60 min groups significantly increased the apoptosis rate of the mouse hippocampus compared with the hypoxia group at corresponding time points(P<0.05).CONCLUSION NaCN can exacerbate hypoxia induced decrease in cerebral blood flow,oxidative stress in brain tissue,and neuro-nal apoptosis in mice,thereby reducing oxygen consumption in closed hypoxic tanks and prolonging their survival time.The mechanism is related to reduced utility of cell oxygen,delaying CO2 accumulation and increasing free oxygen in vivo.