Objective To explore the pathological changes of the livers from hepatic failure (HF)patients and its association with clinical disease stages.Methods Thirty-nine patients with liver failure caused by HBV infections were investigated,and none accompanied with hepatocellular carci- noma.The sections of tissue were taken from the liver after liver transplantation and stained with he- matoxylin eosin(H&E)or RT(reticular fiber)staining.The pathological features were analyzed and compared between the clinical and pathological diagnosis.Results 1.The range and the grade of the pathological changes were all well-proportioned in the whole liver but quite asymmetrical in the same spicemen.2.4 cases with clinical diagnosis of cirrhosis(active stage)were in accordance with the pathological diagnosis.Only 17 in 35cases can be pathologically diagnosed as chronic severe hepatitis (SH),while the other 18 cases were pathologically diagnosed as cirrhosis(active stage).Conclu- sion There were a great inconsistency between the clinical and pathological diagnosis.

Objective To explore the safety of the mobilization of peripheral blood stem cells(PBSC)by granulocyte colony-stimulating factor(G-CSF)in elderly donors.Methods 28 peripheral arteriosclerotic occlusive disease(PAOD)elderly patients(aged≥60 years),and 29 healthy sibling young/adult donors(aged＜60 years)for peripheral allogenic stem cell transplantation were included.Blood samples were collected immediately before starting G-CSF and prior to PBSC collection to analyze the following parameters:the WBC counts,fibrinogen(FIB),D-dimer (D-D),thrombin antithrombin complex(TAT),antithrombin(AT)and yon Willebrand factor antigen(vWF:Ag). Results It had a very significant increase in D-D and vWF:Ag and a very significant decrease of AT(P＜0.01),af- ter mobilization by G-CSF,and a increase in FIB and TAT were also observed(P＜0.05,P＜0.01)in elderly group.In the young/aduh group,the increase in FIB was significant(P＜0.05).The elevating extent of D-D and TAT after G-CSF administration was significantly higher in elderly group than that in young/adult group(P＜0.05).Compared to young/adult group,there was a significant increase in thrombotic events and cerebrovascular ac- cident(P＜0.05).Conclusion In PBCS donorsreceiving G-CSF it reveals activation of both coagulation and en- dothelial cells and inhibition of anticoagulant system that can favor the developing of thrombotic events,which is more remarkable in elderly donors.Therefore a careful monitoring for coagulation system should be considered in those elderly cases.

OBJECTIVETo evaluate the effectiveness of locking plate external fixator in treating middle and distal tibial fractures.

METHODSFrom January 2010 to January 2013,18 patients suffered from middle and distal tibial fractures were treated by locking plate external fixator,including 11 males and 7 females, with an average age of 53.5 (ranged from 13 to 80) years old,the course of disease ranged from 2 h to 3 d. According to AO classification, 4 cases were type A,11 cases were type B and 3 were type C. Among them,6 patients were open fracture, including 2 cases with type I, 3 cases with type II and 1 case with type III, according to Gustilo classification), 12 patients were close fracture. Operation time, postoperative complications were observed, and Johner-Wruhs scoring were used to evaluate clinical outcomes.

RESULTSAll patients were followed up from 6 to 15 (meaned 11) months. Two cases occurred skin necrosis (1 case occurred bone exposure), 2 cases occurred delayed union (all were open fracture), and 1 case occurred nail infection. No screw loosening or broken occurred. According to Johner-Wruhs scoring, 10 cases obtained excellent result,6 cases good,and 2 cases fine.

CONCLUSIONLocking plate external fixator for the treatment of middle and distal tibial fractures, which has advantages of lessen damage, shorter operative time, less complications and rapid functional recovery, is one of good choice.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bone Plates ; utilization ; External Fixators ; utilization ; Female ; Fracture Fixation ; instrumentation ; methods ; Humans ; Male ; Middle Aged ; Tibial Fractures ; surgery ; Treatment Outcome ; Young Adult

Objective To describe the clinical features of venous diethylene glycol poisoning and to identify factors correlating with such kind of poisoning.Methods Retrospective chart review was performed to analyze the epidemiology,clinical presentation,hepatorenal functions,bemodynam- ics and pathological characteristics of 64 patients with severe liver diseases who received intravenous diethylene glycol.Comparative analyses of correlating factors and causes of poisoning were based on the presence or absence of poisoning.Results Fifteen cases of poisoning were reported.After a 5 day incubation period,all poisoned patients displayed acute renal failure and 11 cases with digestive tract symptoms and(or) symptom exacerbations were noted.Neurological system impairment was observed in 10 cases after 2 weeks.Metabolic acidosis developed in 13 cases.Poisoned patients exhibited signif- icantly lower red blood celI(RBC)[(2.32?0.76)?10~(12)/L],hemoglobin(Hb) [(79.5?23.6)g/L] value and higher white blood cell(WBC)[(9.78?3.75)?10~9/L] count.Renal biopsy of poisoned patients revealed acute tubular necrosis and interstitial nephritis.Twelve poisoned patients died.Sig nifieant differences were found between groups regarding preexisting severe hepatitis,ascites,renal disease and diuretic therapy.Prior to diethylene glycol injections,mean values of neutrophil,blood urea nitrogen(BUN),creatinine(Cr) and calcium and phosphorousions differed significantly between groups.Conclusions Features of venous diethylene glycol poisoning include oliguric acute renal fail- ure,metabolic acidosis,digestive symptoms,nervous system impairment and a high probability of anemia and WBC proliferation.Mortality is high.Correlative factors include preexisting severe liver disease,renal disease and infection.

AIM To observe the curative effects of Modified Buzhong Yiqi Decoction (Codonopsis Radix,Astragali Radix,Atractylodis macrocephalae Rhizoma,ect.) on perioperative period of intertrochanteric fractures in elderly patients with Qi deficiency and blood stasis type.METHODS One hundred and twenty cases of elderly intertrochanteric fractures were randomly divided into treatment group and control group,60 cases in each group.All patients underwent closed fracture reduction and proximal femoral nail anti-rotation (PFNA) fixation.During the perioperative period,the control group only received conventional treatment,while the treatment group received conventional treatment and Modified Buzhong Yiqi Decoction once a day.The levels of interleukin-6 (IL-6),interleukin-10 (IL-10) and tumor necrosis factor (TNF-α) of both groups in preoperative and in the 1st day,the 7th day,the 14th day of postoperative were observed and compared.The complications and the curative effects according to the Harris score also were observed and compared between the two groups.RESULTS In preoperative,there was no significant difference in levels of IL-6,IL-10 and TNF-α between the two groups (P ＞ 0.05).The levels of IL-6 and TNF-α in the 1 st day of postoperative in the control group were significantly higher than those in the treatment group (P ＜0.05,P ＜0.01).And the levels of IL-10 were similar between the two groups (P ＞0.05).In addition,in the 7th day and the 14th day of postoperative,the levels of IL-6,IL-10 and TNF-α in both groups were all decreased,moreover,the descent degree in treatment group was more significant than those in the control group (P ＜ 0.05,P ＜ 0.01).The treatment group had the lower incidence of complications and the higher curative effects than those in the control group,both differences were statistically significant (P ＜0.05).CONCLUSION Modified Buzhong Yiqi Decoction applying to the perioperative period of femoral intertrochanteric fractures in elderly patients with Qi deficiency and blood stasis,can effectively reduce the perioperative inflammatory response,reduce the incidence of postoperative complications,and improve the curative effects.So it has a positive effect on the rehabilitation for the traumatic patients and is worthy of clinical promotion.

To increase the in vivo half-life of human CNTF mutein AX15 (R13K), HSA-AX15 (R13K) fusion protein was constructed by the fusion of the C-terminus of HSA to the N-terminus of AX15 (R13K) via an 11 amino acids linker. HSA-AX15 (R13K) fusion protein was purified to homogeneity by cation exchange chromatography, reverse phase chromatography and gel filtration after expressed in pichia pastoris. TF-1 cell survival bioassay showed the biological activity of AX15 (R13K) was not affected by the fusion to HSA. It was demonstrated that tertian injection of 4.8 mg/kg HSA-AX15 (R13K) fusion protein could produce more potent anti-obesity effects on KM mice than daily injection of 1.6 mg/kg AX15 (R13K). The long-acting form of hCNTF variant has the potential to reduce discomfort by requiring fewer injections and to minimize the side-effects by decreasing the dosage and fluctuation of plasma concentration, and thus has superior clinical application.
Animals ; Ciliary Neurotrophic Factor ; genetics ; Humans ; Mice ; Mutant Proteins ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification ; Serum Albumin ; genetics

AX15 is a mutein of naturally occurring human ciliary neurophic factor (hCNTF), with improved biological activity, stability and solubility. AX15 is susceptible to protease degradation when expressed in Pichia pastoris. Amino acid sequencing revealed the degradation was occurred behind position 12 and 13 amino acid residues, which constitute a dibasic site, RR. Based on the substrate specificity of KEX2, a KEX2 resistant mutein of AX15-AX15 (R13K) was constructed, in which RR was replaced by RK. It was demonstrated that the stability of AX15 (R13K) improved significantly, as no degradation was detected even after 120 hours of induction. AX15 (R13K) was purified to homogeneity by ultrafiltration and gel filtration. TF-1 cell survival bioassay showed AX15 (R13K) had equivalent specific activity to AX15. The protease resistant mutein of AX15 may have greater in vivo stability and thus have superior therapeutic potential.
Ciliary Neurotrophic Factor ; biosynthesis ; genetics ; Genetic Vectors ; Humans ; Mutant Proteins ; biosynthesis ; genetics ; Mutation ; Peptide Hydrolases ; chemistry ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification

OBJECTIVETo investigate the protective effect of histone acetylation against hypoxic-ischemic cortical injury in neonatal rats.

METHODSA total of 90 neonatal rats aged 3 days were divided into three groups: sham-operation, cortical injury model, and sodium butyrate (a histone deacetylase inhibitor) treatment. The rats in the model and the sodium butyrate treatment groups were intraperitoneally injected with lipopolysaccharide (0.05 mg/kg), and then right common carotid artery ligation was performed 2 hours later and the rats were put in a hypoxic chamber (oxygen concentration 6.5%) for 90 minutes. The rats in the sham-operation group were intraperitoneally injected with normal saline and the right common carotid artery was only separated and exposed without ligation or hypoxic treatment. The rats in the sodium butyrate treatment group were intraperitoneally injected with sodium butyrate (300 mg/kg) immediately after establishment of the cortical injury model once a day for 7 days. Those in the sham-operation and the model groups were injected with the same volume of normal saline. At 7 days after establishment of the model, Western blot was used to measure the protein expression of histone H3 (HH3), acetylated histone H3 (AH3), B-cell lymphoma/leukemia-2 (Bcl-2), Bcl-2-associated X protein (BAX), cleaved caspase-3 (CC3), and brain-derived neurotrophic factor (BDNF). Immunofluorescence assay was used to measure the expression of 5-bromo-2'-deoxyuridine (BrdU) as the cortex cell proliferation index.

RESULTSThe sodium butyrate treatment group had a significantly lower HH3/AH3 ratio than the model group (P<0.05), which suggested that the sodium butyrate treatment group had increased acetylation of HH3. Compared with the model group, the sodium butyrate treatment group had a significant increase in Bcl-2/Bax ratio, a significant reduction in CC3 expression, and a significant increase in BDNF expression (P<0.05). The sodium butyrate treatment group had a significant increase in the number of BrdU-positive cells in the cortex compared with the model group (P<0.05), and BrdU was mainly expressed in the neurons.

CONCLUSIONSIncreased histone acetylation may protect neonatal rats against cortical injury by reducing apoptosis and promoting regeneration of neurons. The mechanism may be associated with increased expression of BDNF.

Acetylation ; Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Brain-Derived Neurotrophic Factor ; analysis ; Butyric Acid ; therapeutic use ; Cerebral Cortex ; pathology ; Female ; Histones ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

Objective:To investigate the effect of sesamin on mast cell activation and its inflammatory mediator release,as well as its possible mechanisms of action.Methods:HCM-1 cells were activation by stimulation with 10 μg/ml anti-DNP IgE for 6 h and challenge with 100 ng/ml DNP-HAS for 10 min.Sesamin was administration at the concentration of 25,50 and 100 μg/L prior to DNP-HAS challenge,subsequently the effect of sesamin on mast cell degranulation was investigated by light microscope,and histamine release and expression of cytokines such as TNF-α IL-6,IL-1β,IL-8 of mast cells after sesamin treatment were investigated by ELISA.Western blot was used to determine the effect of sesamin on FcεRI downstream signaling including Lyn,Syk and PKCα activation,and IκBα phosphorylation and NF-κB activation.Results:DNP-HAS significantly increased mast cell degranulation,histamine release and those cytokines expression,enhanced Lyn,Syk,PKCα,IκBα phosphorylation and NF-κB activation(P<0.05). Sesamin(50,100 μg/L) significantly decreased mast cell degranulation,histamine release and cytokines expression (TNF-α,IL-4,IL-1β,and IL-8),reduced activity of Lyn,Syk,kinases and PKCα and IκBα phosphorylation,and inhibited NF-κB activation(P<0.05).Conclusion: Sesamin suppresses mast cell activation and inflammatory mediators release through inhibition of PKCα/NF-κB signaling pathway.

Apolipoprotein A-I-Milano(AIM), a natural variant, not only inhibits the initiation and progression of atherosclerosis, but also makes the preexisting atherosclerotic lesions regress. AIM gene, at which N-terminal codens were optimized, was subcloned into the expression vector of pET22b. Recombiant plasmids were transformed into E. coli strain BL21 (DE3) and induced with IPTG. The expressed apoliprotein A-I-Milano was soluble in E. coli and was about 38% of total cell lysate. Purified by Butyl Sepharose 4F. F hydrophobic chromatography and Q Sepharose H.P. anion exchange chromatography, followed by ultrafiltration with Vivaspin 20 (30 000MW), AIM monomer was obtained in a purity of more than 95%. Activity assay of binding of AIM monomer to lipid indicates that association of AIM monomer with DMPC is slower than normal apoA-I but DMPC number associated by AIM monomer is more than by apoA-I. This results will be important for studying structure, function of AIM, specially clinical application.
Apolipoprotein A-I ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Humans ; Mutant Proteins ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Solubility