Objective To investigate the differentiation of mesenchymal stem cells(rMSCs) of young rat into neuron-like cells with Ligustrazin hydrochloride. Methods rMSCs were separated from femurs marrow flushed out with DMEM(low glucose) by using a needle and syringe, then planted in plastic culture flask. Through expanded to 5 passages, rMSCs were induced to differentiate into neuron-like cells with Ligustrazin hydrochloride. Anti-neurofilament(NF-M), nestin, neuron-specific enolase(NSE), MAP-2,GAP-43 and glial fibrillary acidic protein(GFAP) antibodies were detected by immunohistochemistry. Results rMSCs were comprised a single phenotypic population and displayed a fibroblast-like morphology after 5 passage in culture. With 10*!?g/L bFGF pre-induce for 24*!h, then the medium was replaced with induction medium containing Ligustrazin hydrochloride. The induced-rMSCs exhibited neuronal morphological characteristics from the first half an hour to 5*!h. The neuron-like cells expressed NF-M, NSE, MAP-2,GAP-43 and nestin positive, but didn't express glial astrocyte marker GFAP.Conclusion rMSCs can be induced to differentiate into neuron like cells with Ligustrazin hydrochloride in vitro.

Adolescent ; Adult ; Child ; Disasters ; Earthquakes ; Female ; Humans ; Male ; Middle Aged ; Sunburn ; Young Adult

Objective:To investigate the curative effects and mechanism of Shuxuetong injection (疏血通注射液) on treating coronary heart disease(CHD).Methods:The treated group (n=20) based on conventional therapy was treated with Shuxuetong injection infused intravenously,meanwhile the control group treated with conventional therapy only.Before and after therapy the changes in concentrations of plasma endothelin(ET),thromboxin B2(TXB2),6ketoprostaglandin F1α(6ketoPGF1α) and blood rheology between two groups were compared.Results:After therapy the clinical symptoms were convalescent,the ET and TXB2 reduced,the 6ketoPGF1α increased,and the parameters of blood rheology improved.Conclusions:Shuxuetong injection is able to dilate vessels,increase blood flow volume,anticoagulate,and improve blood rheology so that it is effective drug to treat CHD.

Objective To evaluate the feasibility and safety of transjugular liver biopsy( TJLB) by using the LABS 200 liver access and biopsy set ( Cook Inc, USA) .Methods Five minipigs were operated though TJLB puncture under the imaging guidance.The liver biopsies were analyzed by histological examination.Results Technical success of TJLB was achieved in all the 5 minipigs.No procedure-related complications occurred, and sufficient amount of specimen for histological examination was obtained in all cases.Conclusions Our preliminary results indicate that transjugular liver biopsy with the use of Cook LABS 200 liver access and biopsy set is clinically safe and feasible, and provide technical support for its clinical application.

Objective To determinted whether GM1 had a protective effect on injury induced by serum-deprivation and the possible mechanism in PC12 cells. Methods The viability of PC12 cells was quantified by MTT after serum-deprivation.The number of apoptotic cells and necrotic cells were determined by Hoechst 33258/PI staining.And the change of PKC protein expression on PC12 cells' membrane and cytosols was detected by Western blotting. Results 1.The viability of PC12 cells decreased after serum-deprivation and the serum-deprivation for 24 hours was chosen as an injury model in this research.Most of the PC12 cells presented apoptosis 24 hours after serum-deprivation.In addition,the PC12 cells' cytosols PKC protein decreased,while the PC12 cells' membrane PKC protein increased significantly,and this result suggested PKC's translocation to membrane and its activation.2.The viability of PC12 cells preincubated with GM1 in high concentrations(10,1,0.1?mol/L) increased significantly and GM1 protected PC12 cells from apoptosis after serum-deprived injury.GM1 reduced the damage of serum-deprivation on PC12 cells and inhibited PKC protein translocation after injury.3.The repair function of GM1 was effective to neuronal resume after serum-deprived injury.Conclusion Neuroprotective effects of GM1 on serum-deprived injury may be partly mediated through the regulation of PKC pathways and it is helpful for the recovery after injury.

ObjectiveA HPLC-FLD method was developed to determine the levels of serum AAA in CRIpatients, and to studythe variationof serum AAAinCRI patientsanditsclinical significances. MethodsSerumsampleswerecollected from100healthcontrolsand 80CRI patients. According to 2002 National Kidney Foundation (NKF) staging diagnosis, CRI patients included 4 of stage 2, 12 of stage 3, 12 of stage 4, 52 of stage 5. According to pathogenesis, CRI patients were also divided into 3 groups :chronic nephritis group ( n = 32), DM group ( n = 36), hypertension group ( n = 12 ).Serums were deproteinized by equal volume of 5％ (v/v) PCA and supernate were analyzed direcdy. External standard method was used as quantitative method. The analytical column was Megres C18. 10％ acetonitrile in water was used as mobile phase. Flow rote was 1.0 ml/min. The wavelengths of fluorescence excitation and emission were changed with specific time. The levels of Tyr, Phe and Trp in CRI groups, different CKD stages and different pathogenesis were compared with healthy control groups to evaluated the sensitivity and specificity of serum AAA for CRI diagnosis. ResultsThe linear ranges of the method were 0. 550 -275.000, 3. 050 - 1220. 000 and 0. 049 -49. 000 pμmol/L for Tyr, Phe and Trp, respectively. The limit of detection (LOD) was 0.014 μmol/L for Tyr, 0.500μmol/L for Phe, and 0.005 μmol/L for Trp. The average recovery was 100. 9％, 101.3％ and 98. 5％ for Tyr, Phe and Trp, respectively. Intra-day CVwas 3. 18％ -4. 20％ ( mean was 3. 13％ )and inter-day CV was 3. 18％ -4. 20％ ( mean was 3. 58％ ). The concentration of serum AAA, Tyr and Trp and the ratio of Tyr/Phe in CRI patients were( 135.74 ±23.23 )μmol/L, (52.27 +8.25) μmol/L, (21.49 ±4.25) μmol/L and[0.87(0.68 - 1.05)]μmol/L. which were lower than that in healthy groups (t value was -14. 709, 4.452, 22. 100, U value was 266.000,respectively, P＜0. 05). The concentration of serum AAA, Tyr and Trp and the ratio of Tyr/Phe in healthy groups were ( 174. 47 ± 11.57 ) μmol/L, ( 63.53 ± 4. 68 ) μmol/L, (44. 22 ± 3. 67 ) μmol/L and[0. 97(0. 94 - 1.00)]μmoL/L. There were no statistically significant difference between the different stage of CRI. Compared with the concentration of Tyr, Phe and Trp among chronic nephritis group, DM group,hypertension group, the concentration of Tyr had no significant changes among these three kinds of diseases (P ＞ 0. 05 ). The concentration of Phe had significant changes between Chronic nephritis group and DM group, Chronic nephritis group and hypertension group ( U = 395.00, 114. 00, P ＜ 0. 05 ) ; the concentration of Trp haad significant changes between Chronic nephritis group and DM group ( U = 349.00, P ＜ 0. 05 ).The diagnostic sensitivity and specificity of serum AAA for CRI were 90％ (72/80) and 100. 0％ (100/100).ConclusionsThe method of high-performance liquid chromatography with fluorescence detection ( HPLC-FLD) is simple, rapid, sensitive and specific. Simultaneous determination of serum AAA was benefit to the diagnosis and evaluation of CRI patients.

Objective:This study aimed to detect the expression of annexin A1 in human non-small cell lung cancer (NSCLC) and to explore its clinical significance. Methods:We detected the expression levels of annexin A1 in NSCLC and the same patient's dis-tal cancerous tissue (from the edge of the tumor >5 cm) by real-time fluorescence quantitative polymerase chain reaction (real-time PCR), Western blot, and immunohistochemical methods, and then analyzed their correlation with clinical pathological parameters. Re-sults: Real-time PCR showed that the expression of annexin A1 mRNA in NSCLC was higher than that of distal cancerous tissue (0.574±1.403 vs. 0.240±0.893, t=2.060, P=0.045). Moreover, the expression of annexin A1 in NSCLC was associated with the degree of differentiation, lymph node metastasis, and TNM staging (P<0.05), but independent of gender, age, smoking history, tumor size, and histological type (P>0.05). Western blot and immunohistochemistry revealed that the expression of annexin A1 protein in NSCLC was higher than that of distal cancerous tissue. Conclusion:Annexin A1 is highly expressed in the cancer tissues of patients with NSCLC, which may be correlated with the occurrence and development of tumor and its invasion and metastasis.

ObjectiveTo observe the preventive effect of type Ⅱ collagen on experimental rat articular cartilage damage induced by T-2 toxin,to explore molecular biomarkers of articular cartilage damage and repair,and to provide a theoretical basis for control of articular cartilage damage.MethodsEighty Wistar rats were randomly divided into 4 groups according to their body weights:negative control,positive control,high-dose intervention,and low-dose intervention groups,20 rats in each group.Animals in negative control group were fed with standard rat chow,and animals in other three groups were fed with T-2-toxin-contaminated chow( 100 ng/kgfeed).Animals in negative and positive control groups drank distilled water,animals in high-dose intervention and low-dose intervention groups drank water containing type Ⅱ collagen(0.5,5.0 g/L,respectively).These rats were sacrificed after 3 and 5 months,respectively,and bilateral knee joints were collected.Histopathologic changes in hyaline cartilage were examined by light microscope,serum levels of type Ⅱ collagen carboxyl terminal peptide (CTX-Ⅱ ),cartilage oligomeric matrix protein (COMP) and urinary deoxypyridinoline (DPD) were determined by enzyme-linked immunosorbent assay(ELISA).ResultsHE staining showed,that the positive control articular chondrocytes were disarranged,deformated,degenerated,with necrosis and extensive areas of chondrocyte loss;but the two intervention groups only showed fibril formation and swelling and surface cartilage cells became round,flat cartilage cells decreased in number,and cartilage cells clustered and so on early pathological changes of osteoarthritis.At the ends of 3 month and 5 month experiment,the levels of serum CTX- Ⅱ in different groups were,negative control[(18.77 ± 4.61),(25.07 ± 9.17)μg/L],high-dose intervention[ (21.11 ± 5.02),(33.20 ± 9.74)μg/L ],low-dose intervention [ ( 19.87 ± 4.53 ),( 29.73 ± 9.32 ) μg/L ] and positive control [ ( 24.43 ± 5.23 ),( 39.17 ±10.49 ) μg/L ] ; the levels of serum COMP were,negative control group [ (5.43 ± 2.75 ),( 6.38 ± 2.23 ) μg/L ],highdose intervention group[ (17.27 ± 4.77),(20.32 ± 4.74)μg/L],low-dose intervention group[(20.13 ± 5.07),(19.44 ± 4.92)μg/L] and positive control group[ (21.37 ± 4.72),(24.52 ± 4.26)μg/L].At the end of 3 month,compared with negative control group,the level of serum CTX- Ⅱ in other three groups increased,but only positive control group increased significantly(P ＜ 0.05) ; at the end of 5 month,compared with negative control group,the level of serum CTX-Ⅱ in other three groups increased significantly,and the difference was statistically significant (all P ＜ 0.05),and the level of CTX-Ⅱ in the two intervention groups was significantly lower compared with that of positive control group(all P ＜ 0.05).Compared with negative control group,the level of serum COMP in other groups increased significantly at the end of 3 month (all P ＜ 0.05) and only the level of serum COMP in high-dose intervention group was significantly lower compared with that of positive control group(P ＜ 0.05).At the end of 5 month,compared with negative control group,the level of serum COMP in other three groups increased significantly,the difference were statistically significant (all P ＜ 0.05) ; the levels of serum COMP in the two intervention groups were significantly lower than that of positive control group(all P ＜ 0.05).At the ends of 3 month and 5 month,the content of urinary DPD in negative control group were[ (3.47 ± 2.20),(4.14 ± 1.06)μg/L],positive control group[ (4.09 ± 2.48),(4.33 ± 3.43)μg/L],high-dose intervention group[ (3.86 ± 2.31 ),(5.72 ± 3.89)μg/L] and low-dose intervention group[ (3.58 ± 2.77),(4.23 ± 2.90)μg/L].The difference between the 4 groups were not statistically significant (F =2.608,2.436,all P ＞ 0.05).ConclusionsType Ⅱ collagen could effectively reduce the level of serum CTX-Ⅱ and COMP in experimental rats and delay the process of articular cartilage damage induced by T-2 toxin.

ObjectiveTo study the impact of jogging mode on T-2 toxin-induced articular cartilage injury in rats,and to evaluate the role of movement in the development of bone and joint disease.MethodsA hundred Wistar rats were randomly divided into five groups:negative control group(free activities in the cage),positive control group(firee activities in the cage),high-regulation group(regular exercise,the treadmill speed of 24 m/min),lowregulation group (regular exercise,the treadmill speed of 12 m/min) and the random group(random exercise,the treadmill speed of 12 or 24 n/min).The negative control group was fed on commercial grain fodder and other groups were fed on grain fodder contaminated with T-2 toxin.At the end of 5,10 weeks,the histopathological changes of hyaline cartilage were detected by optical microscope,and the level of serum cartilage oligomeric matrix protein (COMP) was determined.ResultsArticular cartilage lesions in each experimental group was evident,presented as cartilage cell degeneration,necrosis,karyopyknosis deeply stained,cells arranged in disorder and cell proliferation,articular dryness,and so on.Compared with the positive control group,the cartilage surface cells of rats in the movement groups showed degeneration,necrosis and loss of cells obviously.The injury in high-regulation group was the most serious than that in other movement groups,with the surface and the middle layer lesions,and a large area of cartilage necrosis,and loss of matrix collagen; cartilage degeneration,polarity disappeared,cell proliferation-based disorder showed in random group.The pathological changes of rat articular cartilage damage worsened with the extension of experimental period.The serum levels of COMP at week 5 in experimental groups were higher than that of both the negative control group and the positive control group,and the difference was statistically significant (F =15.733,P ＜ 0.05 ); compared with negative control group [ (11.55 ± 0.89)μg/L],the COMP levels in high-regulation group,low-regulation group,random group[(13.95 ± 1.23),(14.96 ± 1.29),( 12.99 ± 1.43)μg/L] were significantly higher(all P ＜ 0.05); compared with the positive control group[(12.32 ± 1.38) μg/L],the COMP levels in high-regulation group and low-regulation group were significantly higher(all P ＜ 0.05) ; and compared between the exercise groups,the COMP levels in low-regulatinn group were higher than that of random group(P ＜ 0.05).At week 10,the changes were in the same trend as that of week 5,and the difference between groups was statistically significant (F =6.144,P ＜ 0.05) ; and compared with the negative control group [(10.59 ± 1.93)μg/L],the COMP levels in high-regulation group,low-regulation group,random group [ ( 13.72 ± 2.67 ),( 14.94 ± 1.06 ),( 13.21 ± 1.58 ) μg/L] were significantly higher(all P ＜ 0.05) ; compared with the positive control group[ (11.45 ± 0.12)μg/L],the COMP levels in low-regulation group were significantly higher (P＜0.05); but compared with the exercise groups,the difference were not statistically significant(all P＞0.05).ConclusionsHigh-intensity regular running and irregular intensity running can increase the articular cartilage damage,and injury of articular cartilage by low-intensity treadmill exercise is not significant.

Objective To discuss the feasibility and long-term effect of free latissimus dorsi muscle flap in repair of severe lower extremity injury in children. Methods From July 1999 to June 2004, nine child patients (at age of 6-13 years) with severe lower extremity injury involving soft tissue defects a-round the calf and the foot associated with complex open fractures, bare dislocation, and injury of the nerve, tendon and artery were repaired with free latissimus donsi flap, with flap area ranging from 30 cm ×12 cm to 10 cm × 5 cm. Results All the latissimus dorsi flaps survived, with success rate of 100%. A follow-up for 4-9 years showed that the flap had sound shape and function and normal blood supply, without significant influence on donor area. Conclusion Latissimus dorsi flap has advantages of constant anatomical site, abundant blood supply, massive area, strong anti-infection ability and less in-fluence on donor area and hence is an ideal method for repairing severe lower extremity injury in children.