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.

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.

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 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.

Background and purpose: MORC2 (microrchidia family CW-type zinc finger 2, MORC2) is a newly identified chromatin remodeling protein that plays key roles in DNA-based biological processes including gene transcription and DNA damage repair. However, its functional role in breast cancer development and progression re-mains unknown. ALDH1A3 (aldehyde dehydrogenase 1 family member A3), a member of the aldehyde dehydrogenases (ALDH) superfamily, is a putative breast cancer stem cell marker, but its regulatory mechanism in breast cancer is poor-ly characterized. This study aimed to investigate the effects of knockdown of endogenous MORC2 on the expression levels of ALDH1A3 and the breast cancer stem-like phenotype in MCF-7 cells. Methods: Human breast cancer MCF-7 cells were infected with negative control short hairpin RNAs (shNC) and specific shRNAs targeting human MORC2 (shMORC2), followed by selection with puromycin to generate stable MORC2 gene knockdown cell lines. Western blot and real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) were used to examine the protein and mRNA levels of ALDH1A3 in MCF-7 cells stably expressing shNC and shMORC2. Microsphere formation and fluo-rescence-activated cell sorting (FACS) assays were used to analyze the effects of knockdown of MORC2 on the breast cancer stem-like phenotype. Results: Western blot and RTFQ-PCR analyses revealed that the protein and mRNA levels of ALDH1A3 were significantly down-regulated in shMORC2 expressing cells as compared with shNC -transfected control cells. Moreover, mammosphere formation assay showed that knockdown of endogenous MORC2 in MCF-7 cells significantly reduced the ability of cells to form microspheres. Consistently, FACS assays demonstrated that shMORC2-transfected cells had a lower proportion of ALDH-positive stem cells as compared with shNC expressing cells. In contrast, knockdown of MORC2 did not significantly affect the CD44+CD24- stem cell population. Conclusion:MORC2 promotes a breast cancer stem-like phenotype through, at least in part, regulating ALDH1A3 expression.

Objective To study the application of androgens,AMH for female infertility diagnosis value.Methods Used chemiluminescence to detect androgen testosterone (To),androstenedione (AND),17 (HS)To hydrogen sulfate therapy (17HS),sex hormone binding globulin (SHG)and resistance To seedling le’s hormone (anti-Mullerian hormone,AMH)of 258 cases of patients with female infertility.According to the reason of infertility,female infertility patients were divided into observation group (158 cases of endocrine infertility)and control group (100 cases of tubal factor infertility)and two groups of data had statistical analysis with t test.Used Pearman’s correlation method to analyse the relationship between serum AMH level and AND,SHG in patients wirh female infertility.and used ROC curve to evaluate efficiency of AND and AMH to the diagnosis of female infertility.Results ①The indicators To observation group AND control group,AND,AMH and SHG were (1.25±0.41 vs 0.25±0.15)nmol/L,(4.9±0.62 vs 1.80±0.51)nmol/L,(13.6±3.5 vs 6.4±1.81)ng/ml and (64.2±32.1 vs 89.3±30.2)nmol/L,respectively.Compared with the control group,observation group To,AND and AMH were significantly higher than the control group (t=13.02,11.36,9.35,P values<0.01),but SHG was significantly low-wer than thecontrol group(t=7.35,P<0.01).②Between the biology to produce ets (AMH:7.63~10.1 ng/ml,AND:0.3~3.3 ng/ml,17 HS:18~144μg/dl,SHG:80~560 nmol/L)as the standard,in the observation group:17 HS increased 17.7%,AND increased 72.2%,AMH increased 87.9% and SHG 51.2% reduction.③AMH level and the AND existed positive correlation (r=0.579,P<0.05),negatively correlated with SHG (r=0.763,P<0.05).④AMH,AND and SHG diagnosis of infertility area under the ROC curve (AUC),were 0.921,0.863 and 0.736 respectively,best cutoff value were11.26 ng/ml,4.62 nmol/L and 32.62 ng/ml respectively,and sensitivity of 89.7%,72.9% and 59.6%.Specific degrees were 86.2%,86.2% and 75.6% respectively,and accuracy of 87.1%,87.1% and 81.6%.Jointed inspection of AND,AMH and SHG in the diagnosis of infertility,the sensitivity of the specific degree were 96.3% and 90.2% respectively.Conclusion It showed that AMH,AND and SHG have diagnostic value of internal secretory infertility with ROC curve analysis.De-tection of combined AMH,AND and SHG is more meaningful to the early diagnosis and treatment of infertility.