Objective To investigate the changes of extracellular histones during the course of acute liver failure in mice as well as its therapeutic potential.Methods WT mice (C57BL/6) were randomly (random number) allocated to inducing acute liver failure by lethal doses of GalN/LPS injected i.p.Hepatic function,apoptosis of hepatocytes and histological indexes were measured at different intervals following GalN/LPS challenge.The levels of extracellular histones were determined by using ELISA and Western blot methods.Meanwhile,GalN/LPS-treated mice were administered with anti-histone H3 and antihistone H4 neutralized antibodies,respectively.Results Administration of GalN/LPS to mice caused acute liver failure,characterized by significant elevation of plasma ALT levels and massive hepatocyte apoptosis or necrosis.All mice died within 9-12 hours.The levels of nucleosomes and extracellular histones H3 and H4 were increased considerably in a time-dependent manner.The survival rates in GalN/LPS-treated mice were improved remarkably following administration of anti-histone H3 and H4 neutralized antibodies (P =0.037,P =0.025),likely due to the significant inhibition of TNF-production.Conclusions Extracellular histones are an important mediator implicated in the pathogenesis of acute liver failure.Anti-histones show promising potential in the treatment of acute liver failure,which deserves further investigation in the future.

Objective To evaluate the effect of prosthetic rehabilitation and analyze the exercise training program with myoelectric signal for upper limb stumps.Methods Twenty canes with 22 upper limb stumps were treated with exercise for strength training,muscle contraction and control training and the complication of stump were managed.The myoelectrlc signal of upper limb stumps were detected and the stumps were trained with electronic biofeedback software system,including basic signal of biofeedback training and visual biofeedback training.Then the myoelectric prostheses were assembled.Results After prosthetic rehabilitation and myoelectfic signal training,there Wan no significant atrophy of muscle of stumps,muscle strength and range of motion of these twenty limb stumps increased.The amputees could control muscle contraction and grasp,pinch,wrist rotation,elbow extension or flexion consciously.Twenty myoelectrie prostheses were assembled.Conclusion The prosthetic rehabilitation and myoelectric signal training of limb stump is important for assembling myoelectric prosthesis.

Objective To investigate the effects of two inhibitors of arachidonic acid metabolic pathway (5-cyclooxygenase blockade MK886 and COX 2 blockade celecoxib) on growth and VEGF mRNA expression of human pancreatic cancer cell SW1990.MethodsPancreatic cancer cells SW1990 were cultured with different concentrations of MK886,celecoxib,MK886 and celecoxib,then the cell proliferation was detected by using CCK-8,BLT1 mRNA,PGE2 mRNA and VEGF mRNA expressions were determined by RTPCR.ResultsAfter 10 μmol/L MK886 or 20 mmol/L celecoxib treatment for 24 h,the growth of SW1990 was greatly suppressed ( 1.80 ±0.06 vs 1.65 ±0.10,2.04 ±0.03 vs 1.86 ±0.02,P ＜0.01 ),and the growth suppression of SW1990 cells was increased accompanying the raised concentration of MK886 or celecoxib.After both MK886 and celecoxib treatment for 12 h,the growth of SW 1990 cells was much obviously suppressed (1.72 ±0.05 vs 1.52 ±0.05,P ＜0.01 ).After celecoxib treatment for 48 h,the BLT1 mRNA,PGE2 mRNA and VEGFmRNA expressions were not significantly changed,but the expressions of PGE2 mRNA were significantly decreased ( P ＜ 0.05 ).After MK886 or MK886 + celecoxib treatment,the expressions of BLT1 mRNA,VEGF mRNA were significantly decreased ( P ＜ 0.05 ),but the expressions of PGE2 mRNA were not significantly changed when compared to control group.ConclusionsTwo metabolic pathways of arachidonic acid have a close relation with occurrence and proliferation of pancreatic cancer,when both of the pathways were blocked,the proliferation of the pancreatic cancer cell was suppressed obviously.

Objective To establish the reference range of thyroid function from the normal pregnant women in different pregnant period in Hanzhong region.Methods According to the NACB inclusion criteria,collected local resident pregnant women 4 031 cases,from July 2012 to December 2015.With the combination of random sampling,stratified sampling and cluster sampling,thyroid hormone levels were measured by a fully automated chemiluminescence analyzer and its accompanying reagents.All patients were divided into three groups:998 cases in early pregnancy (T1),1 543 cases in mid-pregnancy (T2),1 490 cases in late pregnancy (T3),and 105 cases of non-pregnant women in childbearing age (T0) were selected as control group.Results The levels of thyroid hormones were different among three periods of pregnant women.TSH were 0.25～5.32,0.42～6.26 and 0.61～7.68 mIU/L respectively in the early,middle and late stages.FT3 were 3.54～6.04,3.57～5.94 and 2.93～5.40 pmol/L,respectively.FT4 was 7.11～16.88,6.78～16.94 and 6.03～16.87 pmol/L.Thyroid hormone levels in pregnant women compared with non-pregnant women,there were significant differences (TSH:x2=233.183,P＜0.05,FT4:x2 =388.12,P＜0.05 and FT3:x2 =558.795,P＜0.05).TSH were lower in early pregnant women comparing to non-pregnancy women,and higher in middle-late pregnant women.The change of FT3 and FT4 were consistent,and reduced with the extension of pregnancy comparing to non-pregnancy women.Conclusion The level of thyroid function in pregnant women were different from non-pregnant women.and the normal reference range of local pregnant specific thyroid hormone should be established.

Objective To analyze the role of a key intracellular signaling molecule GSK3β in hepatocyte apoptosis induced by endoplasmic reticulum stress (ERS).Methods Using mouse hepatoma cell lines(Hepa 1) as cell apoptosis model triggered by tunicamycin,an endoplasmic reticulum stress inducer.One hour before Hepa 1 apoptosis induced by tunicamycin,SB216763 specifically inhibited the activity of GSK3β.Living cells/apoptotic cells were detected using acetoxymethyl (AM)/propidium iodide (PI) staining; Furthermore,the measurement of lactate dehydrogenase(LDH) of cell culture supernatant to evaluate the apoptosis.We detect p-GSK3β,GSK3β,the ERS-related protein(GRP78,CHOP and caspase-12) and caspase-3,cleaved caspase-3 protein expression using Western blot.Results Endoplasmic reticulum stress induced by tunicamycin promotes GSK3β activity; Inhibition of GSK3β activity alleviates endoplasmic reticulum stress:the expression of GRP78,CHOP and caspase-12 expression are inhibited.At the same time,GSK3β activity inhibition significantly reduced the endoplasmic reticulum stress-induced apoptosis:compared to cell apoptosis model group,the intervention group of SB216763 showed that the level of LDH decreased significantly,and PI staining of apoptotic cells was also significant reduction.Western blot results showed that the inhibition of GSK 3 β activity reduced reactive cleaved caspase-3 protein.Conclusion GSK3β is an important signaling molecule in the apoptosis pathway induced by endoplasmic reticulum stress ;Endoplasmic reticulum stress promotes hepatocyte apoptosis by mediating GSK3β.

Objective:To analyze the clinical prognostic factors of liposarcoma on the extremities and trunk, as well as to retrospectively analyze the effect of adjuvant radiotherapy and chemotherapy on liposarcoma of the extremities. Methods:Patients with liposarcoma of the extremities treated in our hospital from July 1, 2007 to December 31, 2012 were followed up. The relationship of clinical prognostic factors with gender, age, location, depth, and size of the tumors, as well as the histological grade and admission status, were statistically analyzed. The effects of adjuvant radiotherapy or chemotherapy on overall survival (OS) and disease free survival (DFS) were evaluated. Results:A total of 82 patients with extremity liposarcoma received surgery-based comprehensive treatment in our hospital. Of the total patients, 73 received a 24-month to 88-month satisfied follow-up;the median follow-up time was 47 months. The OS rate was 83.6%(61/73), and the DFS rate was 68.5%(50/73). Multivariate Cox regression analysis showed that the tumor location, histological grade, and admission status were the independent correlative factors influencing DFS, and the age and pathologic grading were the independent correlative factors influencing the OS. Kaplan-Meier survival analysis showed that radiation therapy can significantly improve the DFS and OS of the G2 and G3-grade liposarcoma (DFS:59.1 months vs. 28.4 months, P<0.01;OS:70.8 months vs. 55.1 months, P<0.05). Significant difference was not found in the effect of chemotherapy on OS and DFS. Conclusion:The prognosis of liposarcoma was significantly associated with the pathologic grades and subtypes. Auxiliary radiotherapy could improve the survival and prognosis of G2 and G3 liposarcoma of the extremities, but the role of chemotherapy in treating liposarcoma remained unclear.

OBJECTIVETo study the role of endoplasmic reticulum stress (ERS) in acute liver failure (ALF) using a mouse model of D-Galactosamine/lipopolysaccharide (D-GalN/LPS)-induced ALF.

METHODSThe ALF model was established by administering intraperitoneal (i.p.) injections of D-Ga1N (700 mg/kg) and LPS (10 mug/kg) to six C57BL/6 mice. Three of the modeled mice were also administered 4-phenylbutyrate (4-PBA; 100 mg/kg i.p.) at 6 hours before the onset of ALF and served as the intervention group. Non-modeled mice served as controls. All mice were analyzed by western blotting and qRT-PCR to determine the expression levels of ERS-related proteins in liver tissue. Liver function was assessed by measuring levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum. Extent of injury to the liver tissue was assessed by hematoxylin-eosin staining and histological analysis. qRT-PCR was also used to detect differences in expression of inflammation-related genes, and western blotting was also used to detect differences in expression of the apoptosis related protein Caspase-3.The extent of apoptosis in liver tissue was assessed by TUNEL assay.

RESULTSThe ERS markers GRP78 and GRP94 showed increased expression at both the gene and protein levels which followed progression of ALF. The ERS effector proteins XBP-1, ATF-6 and IRE 1 a involved in the unfolded protein response were activated in the early stages of ALF, and the ERS-induced apoptosis regulators Caspase-12 and CHOP were activated in the late stage of ALF. Inhibition of ERS by 4-PBA intervention protected against injury to liver tissue and function, as evidenced by significantly lower levels of serum ALT and AST and a remarkably decreased extent of histological alterations. Furthermore, the inhibition of ERS suppressed expression of the proinflammatory cytokines TNFa, IL-6 and IL-1 β, and reduced the extent of hepatocyte apoptosis.

CONCLUSIONERS is activated in the mouse model of D-GalN/LPS-induced ALF. Inhibition of ERS may be protective against liver injury and the mechanism of action may involve reductions in inflammatory and apoptotic factors and/or signaling. Therefore, inhibiting ERS may represent a novel therapeutic approach for treating ALF.

Animals ; Apoptosis ; Disease Models, Animal ; Endoplasmic Reticulum Stress ; Galactosamine ; adverse effects ; Lipopolysaccharides ; adverse effects ; Liver Failure, Acute ; chemically induced ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred C57BL

Objective: To investigate the treatment of nasal deformity in patients with unicoronal craniosynostosis. Methods: In patients over 6 months old, the nasal bones were wedge-removed without fixation. The management of all patients with unicoronal craniosynostosis was distraction osteogenesis of pedicled unilateral frontal bone flap. Results: Postoperative extended distance of the frontal bone was 28—41 mm (mean, 35.4 mm). After extension, three-dimensional reconstruction of cranial CT was carried out, which showed that CVAI was 0.8—1.2 (mean, 0.98), tending to normal. After discharge, dynamic cranial braces were put on for 1 year. Postoperatively, the children were followed up for 8—36 months (mean, 28 months). The shape and nasal deformity of all children were improved compared with those before surgery. Conclusions Nasal wedge resection should be used to correct nasal deformity in children over 6 months with unicoronal craniosynostosis.

Objective To investigate the role of Notch signaling pathway to maintain the stem cell-like characteristics of osteosarcoma and its underlying mechanism.Methods Lentiviral NICD1 or Numb-shRNA was transduced into MG63 osteosarcoma cells to activate Notch activity in vitro.The impact of Notch on osteosarcoma stem cells were assessed by the tumor sphere formation assay and flow cytometry analysis of cell surface markers STRO-1/CD117.The expression of stem cell related genes (Sox2,Oct4) were evaluated by Western blot and qPCR.The nude mice were randomly divided into 3 groups:the NICD1 overexpression (NICD-OE) group,the DAPT group and the control (CON) group.The tumor growth was monitored for 8 weeks and the tumor volume and weight were recorded weekly.To investigate whether Notch regulates Eph pathway,Eph pathway related protein EphB,pEphB was measured by Western blot.The impact of ephrinB 1 on NICD overexpression cell were assessed by tumor sphere formation assay.The expression of Sox2 and Oct4 was evaluated by Western blot.Results NICD1 overexpression or Numb-shRNA increased the activity of Notch pathway.The Notch-activated osteosarcoma showed enhanced in vitro tumor spheroid formation capacity,increased Stro-1/CD117double positive ratio,and upregulated expression of Sox2 and Oct4 in vitro.In animal experiments,it was found that activation of Notch pathway promoted tumor formation in vivo and Notch inhibition decreased it.The primary osteosarcoma cells were obtained from mice xenograft treated with DAPT and its tumor sphere formation capacity was significantly reduced.Finally,The Notch pathway inhibits the phosphorylation of EphB,as well as the downstream signal pathway of EphB,but there is no significant change in total EphB.The activation of Eph pathway inhibited Notch induced up-regulation of tumor sphere formation and Sox2 and Oct4 expression.Conclusion Notch signaling pathway maintains the stem cell-like characteristics of osteosarcoma probably by inhibiting the Eph pathway.

Objective To analyze the clinical characteristics and changes of serum tumor markers in lung cancer patients with different smoking status in Hanzhong area. Methods A retrospective analysis was performed on 642 hospitalized lung cancer patients in Hanzhong area from March 2017 to March 2019. According to their smoking status, they were divided into observation group (smoking history, n=404) and control group (no smoking history, n=238). Age, sex, place of residence, basic information of the disease including pathological stage, pathological type, short-term efficacy, survival and serum tumor marker level were analyzed retrospectively. Results The proportion of male in observation group (67.08%) was significantly higher(57.56%) (χ²=5.855,P<0.05). Observed a group of 50 or more patients (80.94%) were significantly higher (73.53%) (χ²=4.824 , P<0.05); There was no significant difference between the two groups (χ²=2.110 , P<0.05). The proportion of adenocarcinoma in the control group (49.16%) was the highest, and that of small cell lung cancer in the observation group (41.34%) was the highest (χ²=15.291, P <0.05). Comparison of pathological stages between the two groups showed that stage IIIB (32.77%) was the highest in the control group, followed by stage IV (23.53%), Stage IIIA (20.59%), Stage II (13.03%) and stage I (10.08%). The observation group had the highest proportion in stage IIIA (35.40%), followed by Stage IIIB (25.00%), stage IV (16.09%), Stage IIIA (16.09%), stage II (15.10%) and stage I (8.42%). The difference between the two groups was statistically significant (χ²=10.817,P<0.05). Before treatment, serum CEA, CA125 and CA199 levels in observation group were significantly higher (P<0.05). After treatment, the levels of serum tumor markers in both groups were significantly decreased (P<0.05); The serum CEA, CA125 and CA199 levels in observation group were significantly higher (P<0.05). After treatment, THE ORR of the observation group (48.76%) was lower than that of the control group (53.78%), but the difference was not statistically significant (χ²=2.051, P>0.05). The 1-year survival rate of the observation group (64.85%) was significantly lower than that of the control group (73.95%) (χ²=5.255, P<0.05). Conclusion Middle-aged and elderly male smokers in Hanzhong area have a high incidence of lung cancer, multiple stage Ⅲ squamous cell carcinoma, and the level of tumor markers in serum is higher than that of non-smokers. The prognosis is not good, so we should encourage patients to quit smoking, which can improve the survival rate of patients.