A gas chromatographic method has been developed for determining glycerin concentrations of both therapeutic and physiological levels in serum. The linear relationship was obtained in the range of 1.6-800 ug/ml and the coefficient variations determined within a day and between days were 2.2% and 2.4% respectively. Recoveries of glycerol from serum were about 87%.

Objective To study the effect of intra-articular injection of dehydroepiandrosterone (DHEA) on the balance of ADAMTS/TIMP-3 system in a rabbit osteoarthritis models.Methods Sixty rabbits were underwent bilateral anterior cruciate ligament transection (ACLT).Rabbits were randomizedn to the following treatment:one knee of each rabbit was treated with 100 μmol/L DHEA dissolved in dimethylsulphoxide (the experimental group) and the other knee was treated under the same schedule using dimethylsulphoxide (the control group) 4 weeks after transection,once a week for eight weeks.Twelve weeks after ACLT,all rabbits were killed after X-ray assessment and the knee joints were evaluated by gross morphology and histology.The concentration of hydroxyproline and glycosaminoglycan in the cartilage were analyzed.The mRNA expression of ADAMTS-4,ADAMTS-5,tissue inhibitor of metalloproteinases-3 (TIMP-3),transforming growth factor (TGF)-β1.Aggrecan and Collagen Ⅱ in the cartilage were analyzed using reverse transcription polymerase chain reaction (RT-PCR).The protein expression of aggrecan ARGxx and Collagen Ⅱ in the cartilage were analyzed by Western blot.Results By Mann-Whitney test,Gross morphologic scores on femoral condyle and tibial plateau in the control group were significantly higher than the experi-mental group (Z=-3.517,P＜0.01 ; Z=-2.518,P＜0.05).By unpaired Student's t test,histological evaluation showed that the grade of cartilage damage in the experimental group [(5.3±1.2) μg/ml] were less severe than that in the control group (10.1 ± 1.3,P＜0.01).The concentration of hydroxyproline [(5.7±0.3,23.6± 1.7) μg/ml] and glycosaminoglycan (30±4) in the experimental group increased significantly when compared with the control group [(4.6±0.5),(18.5±1.4),(24±4) μg/ml,P＜0.01].The mRNA expression of ADAMTS-4 (0.15±0.03)and ADAMTS-5 (0.10±0.04) in the experimental group decreased significantly compared with the control group (0.29±0.08,0.15±0.05; all P＜0.05).The mRNA expression of TIMP-3 (0.85±0.10),TGF-β1(1.2±0.4),Aggrecan (0.87±0.31) and Collagen Ⅱ (2.74±0.59) in the experimental group increased significantly when compared with the control group (0.70±0.13,0.8±0.4,0.49±0.16,2.2±0.5; all P＜0.05).The protein expression of Aggrecan ARGxx (0.53±0.10) in the experimental group decreased significantly when compared with the control group (0.81±0.12,P＜0.01).The protein expression of Collagen Ⅱ (2.3±0.7) in the experimental group increased significantly when compared with the control group (1.7±0.5,P＜0.05).Conclusion DHEA protects against cartilage degradation and inhibits the progression of OA,TGF-β1,Aggrecan and Collagen Ⅱ in cartilage may be the mechanism of the protective effect of DHEA on OA.

Objective To observe the morphologic protection effect of intra-articular injection of osteoprotegerin (OPG)on articular cartilage in a rabbit model of osteoarthritis (OA).Methods Sixty male New Zealand rabbits were randomly divided into 3 groups:OPG group (n=20),sham-operated group (n=20) and PBS group (n=20).In OPG group and PBS group,each rabbit underwent anterior cruciate ligament transection (ACLT) in left knee joint,then 0.1 ml OPG solution or PBS were injected into the left knee for 8 weeks (5 times a week) in OPG group and PBS group,respectively.In sham-operated group,the anterior cruciate ligament was just exposed without transection,and then the incision was closed.All rabbits were sacrificed 12 weeks after operation,and the left knee joints were obtained.The Pelletier score and Mankin score were used to evaluate the macroscopic and microscopic cartilage morphology.Results The score of femoral condyle cartilage and tibial plateau cartilage was 1.80±0.89 and 1.80±0.77 in OPG group,respectively,and 3.10±0.97 and 3.20±0.77 in PBS group.However,there was no statistical difference in Pelletier score between the sham-operated group and the OPG group.Consistent with the macroscopic data,the OPG group has a significantly lower Mankin score involving cartilage structure (2.65±0.88),chondrocyte (1.35±0.71),Safranin O staining (1.83±0.83),tidemark integrity (0.30±0.47) and total score (6.13±1.97) compared with the PBS group (4.52±1.09,1.85±0.63,2.80±0.75,0.65±0.49,and 9.83±1.98,respectively).The OPG group has a significantly higher total Mankin score compared with the sham-operated group (4.80±1.25),however,there were no statistical differences in four subitem scores between the two groups.Moreover,the cartilage thickness in OPG group was 371.84±38.94 μm,255.09±74.82 μm in PBS group,and 404.68±15.97 μm in the sham-operated group,and the differences were statistically significant between three groups.Conclusion OPG has a protective effect on articular cartilage and can slow the progression of OA by reducing the grade of synovitis,decreasing the volume of osteophytes,and suppressing the loss of cartilage thickness.

Objective To investigate effect of taurine on respiration chain enzyme activity of mitochondria 24 hours after severe traumatic brain injury (TBI) in rats.Methods Fifty-six SD rats were divided into sham group,TBI group,taurine treatment group,and taurine prevention group according to the random number table,with 14 rats per group.Fluid percussion brain injury models were used.Via the caudal vein,normal saline was administered to animals in sham and traumatic brain injury groups immediately after injury,while taurine (200 mg/kg)was administered to animals in taurine treatment group after injury and in taurine prevention group 4 days before injury.Brains were harvested 24 hours postinjury for assays of HE staining and electron microscopy.Mitochondrial respiratory chain complex Ⅰ-Ⅴ activities were detected.Results TBI group presented swelling neurocytes,cell loss,karyopyknosis,shortened even vanished process,and inflammation cell infiltration at the edge of necrosis in HE staining.By contrast,morphological improvement was significant in taurine treatment group but only some neurons were intact in taurine prevention group.Swelling mitochondria and broken or vanished mitochondrial crests were seen in TBI group under the electron microscope.However,normal or minor swelling mitochondria was seen in taurine treatment group and cytoplasm slightly porous and absence of mitochondrial crests were seen in taurine prevention group.Activities of complex Ⅰ,Ⅱ and Ⅴ were significant lower in TBI group (32.52±2.41,4.68 ±0.15,2.49 ±0.73) compared to those in sham group (34.03 ±0.46,5.04 ±0.29,3.20±0.68) and in taurine treatment group (33.95±0.85,5.12-±0.23,3.53 ±0.48) (P＜0.05).And complex Ⅰ in taurine prevention group was significantly enhanced as well (34.44 ± 0.36,P ＜ 0.05).Conclusion Taurine may protect the brain tissues and mitochondrial structure from impairment in TBI rats by improving mitochondrial enzymes activity and reducing secondary energy loss.

AIM:To look for a better method to deal with interstitial lung disease,interferon-gamma(IFN-?)combined with methylprednisolone(M-pred)to influence human embryonic lung fibroblast on proliferation,collagen synthesis and the expression of transforming growth factor-?1(TGF-?1)protein and mRNA were investigated.METHODS:Exponentially growing cells were preincubated for 48 h before harvested.The microculture tetrazolium(MTT)assay was used to measure the inhibition ratios of M-pred combined with different concentrations of IFN-?.The expression of proliferation cell nuclear antigen(PCNA)was detected by immunocytochemical analysis.Hydroxyproline kit was adopted to detect collagen synthesis.The expressions of TGF-?1 mRNA and protein were detected respectively by RT-PCR and Western blotting.RESULTS:Methylprednisolone,IFN-? as well as the combination of methylprednisolone and IFN-? inhibited the proliferation of HELF and the expression of PCNA in comparison with control group(P

Objective To investigate the effect of mild hypothermia on neuroprotection and prognosis prediction of rats with traumatic brain injury (TBI) by dynamically monitoring the somatosensory evoked potentials (SEP) and quantitative electroencephalogram (QEEG).Methods Forty healthy adult male SD rats were randomly divided into four groups according to random number table,ie,normal control group (with no intervention),sham operation group (fenestration only,without drilling),TBI group (fluid percussion was used to produce moderate to severe TBI),and mild hypothemia group (ice blanket was used immediately after TBI for continuous physical cooling and rectal temperature was maintained at 32-35℃ and rewarmed to 37℃ 6 hours after the initiation of cooling),with 10 rats per group.Changes of SEP and QEEG in all groups were monitored at 6,24 hours,and 7 days after TBI.Results (1) Compared with TBI group,the latency of SEP waves (P1 and N1) on the injured side in mild hypothemia group began to shorten at 24 hours(P ＜ 0.05) and were close to that in the sham operation group at 7 days.(2) Except for normal control group and sham operation group,QEEG in TBI group showed decrease of α rhythm,increase of reactivity slow waves,and decrease or disappearance of QEEG relative power spectral values at all time points.In mild hypothermia group,the reactivity slow waves were decreased with a small amount of α wave; QEEG relative power spectral values were increased at 24 hours and 7 days (especially at 24 hours),but werc still lower than those in normal control group (P ＜ 0.05).Conclusion Mild hypothermia exerts neuroprotective effect through reducing SEP latency,raising relative power spectral values of QEEG,and improving the nerve conduction and brain electrical activity of the injured side.

During the mission of Peace friendship-2015, between September 2015 and January 2016,the Chinese Navy hospital ship respectively participated in the Harmony-2015joint military exercise in Malaysia and with the local military of Polynesia and Barbados .The article summarized the relevant background of the joint military exercise and the main practices in various stages .This article also suggests that the emergency platform be moved forward , the function of the triage area be expanded, basic life support(BLS),advanced cardiovascular life support (ACLS),and advanced trauma life support(ATLS) training programs be introduced, the training model and the test system be innovated , and realistic traumatic conditions be designed in order to guarantee the success of the joint exercise .

Objective To discuss biomechanical mechanism of reconstructed joint capsule ligament stabilizing hip joint by means of three-dimensional finite element analysis.Methods A finite element model of total hip arthroplasty(THA)including ischiofemoral ligament reconstruction was construtted by using finite element analysis software Unigraphics NX 2.0 and SolidWorks 2006 to simulate seated leg crossing and obtain peak resisting moment and range of motion prior to impingement.Results The current form of finite element model was characterized by large deformation multi-body contact,large interfacial sliding and high elasticity and could clearly reflect real anatomy and biomechanical behavior of ischiofemoral ligaments.Compared with model with only metal,model of ischiofemoral ligament reconstruction could reduce the peak polyethylene stress at the impingement site and at the head egress site by typically 17%and 31%respectively,increase peak resisting moment by nearly 57%and provide 2.29-fold stability.Conclusions As a discrete structure within the posterior capsule of the hip joint,the ischiofemoral ligament may be the most important contributor to the mechanical integrity of the posterior stability structure.The joint capsule ligament must be reconstructed in hip arthroplasty.

Objective To investigate the clinical significance of flow cytometry (FCM) assay in following up of the minimal residual disease (MRD) used for predicting relapse and guiding chemotherapy. Methods The clinical data of 43 acute leukemia patients diagnosed by MIC were collected in our hospital from 2005 July to 2008 June.Bone marrow aspirates were collected from 43 patients with newly diagnosed acute leukemia after induction therapy and during constimulation therapy. The cells with leukemia associated with immunophenotype were investigated using FCM, as immunologic target of MRD. Results MRD were detected earlier in predicting the relapse than those of the traditional bone marrow cells morphology assay by an average of 4-6 months. The results of the MRD following up: MRD was negative at CR in 26 cases, 6 cases relapse, 20 cases of them were kept negative during following up. MRD was positive in 17 cases at CR, 9 cases of them were relapse. 4 cases after intensified chemotherapy the MRD became negative and kept egative for more than one year. The MRD of the 43 cases at CR were divided into 3 groups, MRD less than 1×10-4 group (A group) MRD between 5×10-3 and 1×10-4 group (B group) and MRD above 5×10-3 group(C group). By chi square test. There was no statistical significance between A group and B group, but there was tatistical significance between B group and C group (P=0.02). Conclusion The application of FCM in detecting MRD has important clinical significance in predicting relapse and guiding chemotherapy.

Objective To analyze the conding region of hantanvirus S gene and predict the structure of nucleoprotein for diagnostic antigen study.Methods RT-PCR was used to amplify the S gene of hantanvirus Hunan03 strain after designing specific primers.The amplification product was cloned into pGM-T vector and then the recombinant vector was transformed into E.coli TOP10,gene sequencing was carried out after blue-white selection and PCR screening for positive clones.The database of NCBI and Swiss-Prot/TrEMBL were used to predict and analyze the structure,biological characteristics and protein structures of S gene.Results The amplification product was about 1290 bp,the pGM-T/S vector was constructed and successfully sequenced,the whole length of the open reading frame (ORF) was composed of 1290 nucleotide residues,among them the GC content was 44.11％ and the AT content was 55.89％,it was composed of 429 amino acids (20 kinds),the accession number of the sequence submitted to GenBank was JN712306,its homology of nucleotides to the 76-118 strain was 83％ and the homology of amino acids was 98％,ten nonspecific variation sites were found.The grand average of hydropathicity was-0.405.There were three transmembrane domains and four non transmembrane domains in the secondary structure of nucleoprotein including 55％ of helix structure,6.1％ of sheet structure and 38.9％ of loop structure.Conclusion The bioinformatics analysis of Hunan03 strain S gene might be important for provide the substructure data to reveal the significance of S gene characteristics on hemorrhagic fever renal syndrome (HFRS) prevention and control.