BACKGROUND:There are numerous studies on bone marrow mesenchymal stem cells(BMSCs)from small animals such as rats and rabbits,but no few reports addressing BMSCs from big animals.OBJECTIVE:To observe in vitro cultured goat BMSCs,and to understand its biological properties.METHODS:A healthy Chinese goat aged ten months was obtained to extract 5 mL fresh bone marrow from the posterior superior iliac spine by puncture following anesthesia.Using the whole bone marrow method,the samples were incubated in a sterile plastic culture flask and added with DMEM/F12 containing 10% fetal bovine serum.Following 80% 90% confluence,cells were digested by trypsin.Cells at passage 3 in logarithmic phase were collected and frozen,and then recovered.Changes in cell morphology were observed using an inverted microscope.Cell growth curve was measured using MTT assay.The potential of osteogenic differentiation was examined utilizing Von Kossa's staining.RESULTS AND CONCLUSION:The primary cultured BMSCs were cultured with adherent growth.The cells were spindle form.Cell morphology following passage 3 was similar,showing long spindle shape.Following freezing and recovery,cell adherence was slower compared with subculture cells,and no significant difference was detected in cell morphology and viability compared with subculture cells.Growth cycle was similar in passage 3-passage 5 cells.BMSCs entered lag phase at days 2 and 3,logarithmic phase at day 3,and platform phase at days 6 and 7,and then growth speed was slow.Goat BMSCs were positive for Von Kossa's stain at 3 weeks following osteogenic induction.Results verified that cultured goat BMSCs showed strong genetic stability and proliferation ability,and differentiated into osteoblasts.

Objective To prepare monoclonal antibody(McAb) against human tissue kallikrein (HK) and develop an ELISA kit allows for the in vitro quantitative determination of human tissue kallikrein in urine. Methods To generate a monoclonal antibody specific for TK, the synthetic TK peptide consisting of 12 amine acids(12P), was fused to keyhole limpet hemocyanin(KLH) and used for immunization. Using hybridoma screening, monoclonal secreting cell lines were identified and used to generate ascites in BALB/c mouse. Antibody was purified by affinity column chromatography. 12% SDS-PAGE and Western blot were used to visualize the purified antibody. This kit employs indirect competitive ELISA technique and BiotinAvidin System. 12P was fused to bovine serum albumin(BSA) and has been pre-coated onto a microplate at first. Standards and samples were added to the appropriate microplate wells with a biotin-conjugated McAb croplate well. A TMB substrate solution is added to each well. The enzyme-substrate reaction is terminating by the addition of a sulphuric acid solution and the color change is measured spectrotometrically at a wavelength of 450 nm. The concentration of tissue kallikrein in the samples is then determined by comparing the O.D. of the samples to the standard curve. Results 8 hybridoma cell lines secreting mAbs special to HK,SDS-PAGE and Western blot demonstrated successful preparing and purification of McAb( 100% ). The linearity of this ELISA kit is demonstrated(r =0. 990). The range of detection of the assay is 0.008 μg/ml to 0. 5 μg/ml. The assay remained stable, with no change in the values measured, over five cycles of freezing and thawing. Conclusion 8 McAbs against HK have been prepared successfully and possess high titer and specificity. The development of an ELISA kit for detecting HK can meet the needs of detection of HK in urine samples.

The clinicopathological data of 173 cases of inflammatory myofibroblastic tumor (IMT)were retrospectively analyzed.Among them 125 were males and 48 females with a mean age of 47.9 y (5-78 y).The lesions of 117 cases were located in lungs,41 cases in eyes,8 in ileocecum,2 in liver,2 in spleen,1 in abdominal wall,1 in maxillary sinus and 1 in face.The average diameter of lesions were 3.5 cm,ranging from 1.0 cm to 7.0 cm.The clinical manifestations were not specific,depending on the locations of tumor.The imaging examinations were helpful for diagnosis.The prcsence of slender-spindled myofibroblasts and proliferation of fibroblast cells,with infiltration of chronic inflammatory cells were the basic histopathological features of the disease.Combined with the histological characteristics and immunohistochemical staining IMT can be differentiated from other spindle cell tumors.Appropriate surgical resection is the main treatment for IMT.

Objective To investigate the effect of platelet-rich plasma (PRP) on starting of wnt3 gene and klotho gene of adipose-derived stem cells (ADSCs) of rabbit.Methods Epididymal adipose tissue stem cells were obtained from New Zealand white rabbits,and the cells identified by morphology and inducing differentiation,and the cells were cultured to the fourth generation,PRP and PPP (platelet-poor plasma) were prepared by traditional centrifugal method from abdominal aortic of rabbit; ADSCs were cultured in culture medium containing PRP (experimental group),PPP (control group) and all medium (blank group) for each 5％ for 24h,48h and 72h.Cells of each group were dissociated and total RNA extracted.Effects of the starting of wnt3 gene and klotho gene were detected by RT-PCR.Results Primary ADSCs of rabbit grew in the way of long spindle swirly.The results of oil red O and alizarin red staining of the ADSCs were positive.Expression of wnt3 gene and klotho gene in the experimental group significantly increased from the results of RT-PCR (P＜0.05).Conclusions PRP can promote proliferation of the ADSCs of rabbit and increase the expression of wnt3 gene and klotho gene significantly.

OBJECTIVETo observe the inhibitory effect of gefitineb on the proliferation and its inducing effect on the apoptosis of mouse I-10 Leydig testicular cancer cells in vitro.

METHODSWe treated I-10 Leydig testicular cancer cells of mice with gefitineb at 0, 1.25, 2.5, 5, 10, 20, and 40 µmol/L. Then we determined the inhibitory effect of gefitineb on the growth of the cells by MTT, detected their early and late apoptosis by Annexin V-FITC/propidium iodide double staining and Hoechst 33258 nuclear staining, respectively, and observed the expressions of apoptosis-related proteins Bcl-2, Bax and caspase 3/9 by Western blot.

RESULTSCompared with the blank control group, gefitineb significantly inhibited the proliferation of the I-10 cells at 10 and 20 µmol/L (P < 0.05). The survival rate of the cells was (32.4 ± 2.8)% (P < 0.01) and their early and late apoptosis rates were (26.7 ± 4.2)% and (59.33 ± 10.2)% in the 40 µmol/L group, significantly different from those in the control (P < 0.05 and P <0.01). In comparison with the blank control group, gefitineb at 10, 20, and 40 µmol/L increased the expression of pro-apoptotic protein Bax by (41.9 ± 7.1), (60.1 ± 9.8), and (69.0 ± 11.3)% (all P < 0.05), decreased that of apoptosis-inhibitory protein Bcl-2 by (50.3 ± 8.9), (63.9 ± 6.9), and (88.7 ± 13.9)% (all P < 0.05), and elevated that of the cleft proteins caspase-3 by (69.0 ± 6.9)% (P < 0.05), (71.5 ± 8.1)% (P < 0.05), and (110.9 ± 14.2)% (P < 0.01) and caspase-9 by (51.8 ± 4.9), (54.7 ± 6.7), and (43.8 ± 11.8)% (all P < 0.05).

CONCLUSIONGefitineb can increase the cytotoxicity of I-10 Leydig testicular cancer cells of mice and induce their apoptosis via the mitochondria-mediated apoptosis signaling pathway.

Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; metabolism ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Proliferation ; drug effects ; Cell Survival ; Leydig Cell Tumor ; drug therapy ; metabolism ; pathology ; Male ; Mice ; Neoplasm Proteins ; metabolism ; Neoplasms, Germ Cell and Embryonal ; drug therapy ; metabolism ; pathology ; Quinazolines ; pharmacology ; Testicular Neoplasms ; drug therapy ; metabolism ; pathology ; bcl-2-Associated X Protein ; metabolism

Adult ; Female ; Humans ; Male ; Middle Aged ; Occupational Health ; statistics & numerical data ; Regression Analysis ; Risk Factors ; Stress, Psychological ; psychology ; Surveys and Questionnaires ; Workplace ; psychology

Aims To study the effects of clenbuterol on anoxia/reoxygenation( A/R) injury in neonatal Wistar rat cardiomyocytes and to explore whether its mecha-nism is related to reperfusion injury salvage kinase ( RISK) or not. Methods The cultured primary neo-natal cardiomyocytes were randomly divided into eight groups: ①normal culture group; ②anoxia/reoxygen-ation( A/R) group;③ clenbuterol ( 1 μmol · L-1 ) +A/R;④ICI118,551(10 μmol·L-1) + clenbuterol ( 1 μmol · L-1 ) + A/R; ⑤Metoprolol ( 10μmol · L-1 ) + clenbuterol(1μmol·L-1 ) + A/R group;⑥Metoprolol ( 10 μmol · L-1 ) + A/R group; ⑦PD98059 ( 20 μmol · L-1 ) + clenbuterol ( 1 μmol · L-1 ) + A/R group;⑧ LY294002(10 μmol·L-1 ) +clenbuterol(1 μmol · L-1 ) + A/R group. Cell via-bility was determined by the conventional MTT reduc-tion assay. The content of LDH in cultured medium was measured with colorimetry. Cardiomyocyte apopto-sis was determined by Hoechst33342 . Intracellular re-active species( ROS) were monitored by the fluorescent DCFH-DA. Total ERK2 and phosphorylated ERK were detected by western blot. Results Compared with A/R group, clenbuterol significantly increased vaibility of cells, reduced LDH release, lowered the rate of apop-tosis and ROS production. When addedβ2 receptor an-tagonist ICI118 , 551 , PI3 K inhibitor LY294002 and ERK inhibitor PD98059 , the effects of clenbuterol a-bove were inhibited; but β1 receptor antagonist Meto-prolol protected the cardiomyocytes from A/R injury, as evidenced by decreased LDH release and increased cell viability. There were no synergistic effects in the combined use of clenbuterol and Metoprolol. Conclu-sion clenbuterol exerts cardioprotective effects against A/R injury by inhibiting oxidative stress and apopto-sis. The protection of clenbuterol is inhibited by ICI118 , 551 , LY294002 and PD98059 . clenbuterol protects cardiomyocytes against A/R injury via RISK pathway by activation of β2 receptor.

?AIM: To retrospectively analyze the surgical strategies and outcome of traumatic lens dislocation.?METHODS: Retrospective study. Clinical data of 105 cases ( 105 eyes ) diagnosed with traumatic lens dislocation from April to June 2014 in our hospital were recruited. According to position of dislocated lens and complicated situations, different surgical approaches were performed, including intracapsular lens extraction, phacoemulsification, vitrectomy through pars plana and lensectomy. Meanwhile, vitreo-retinal or anti-glaucoma surgeries were performed in complicated cases. Preoperative and postoperative LogMar ( Logarithm of the Minimum Angle of Resolution ) visual acuity were compared by paired t-test. Perioperative complications including expulsive choroidal hemorrhages and recurrent retinal detachment were recorded and assessed.?RESULTS: All 105 dislocated lenses were removed completely. Visual acuity of 91 eyes ( 86. 7%) were significantly improved postoperatively. The visual acuity of most patients was 0. 1-0. 3 ( 42 eyes, 40. 0%) and 1 patient’s visual acuity with lens subluxation reached more than 0. 8 postoperatively. Expulsive choroidal hemorrhages occurred in 1 eye intraoperatively and 1 eye postoperatively. Recurrent retinal detachment was observed in 2 eyes postoperatively.? CONCLUSION: According to position of the lens dislocation, personalized surgery strategy is critical for therapy of traumatic lens dislocation. Expulsive choroidal hemorrhage is one of most several complications and should be managed properly.

Objective To evaluate the using of either 225 or 150 microgrammes of mifepristone combined with misoprostol for termination of second-trimester gestation(16～24 weeks).Methods 180 women requesting voluntary induced abortion during gestation 16～24 weeks were randomised to three groups,group 1:oral mifepris- tone 225rag,group 2:oral mifepristone 150mg,and group 3:injected 100rag rivanot by amniocentestis.The total suc- cess rate,once success rate,the interval of having-medicine to uterine-constraction,the volume of bleeding within 2 hours after labour and cervical laceration rate were observed.Results The once success rate of induced labour in group 1 was higher than that in group 2 and group 3(P

·Age-related macular degeneration (AMD) and retinitis pigmentosa (RP) are common outer retinal degenerative problems, which are also the predominant causes of most blinding retinal diseases. Retinal prosthesis is a promising solution for such photoreceptor degeneration diseases.Most of current concepts for a retinal prosthesis are based on neuronal electrical stimulation. In the past twenty years, retinal prosthesis has been developed in two different directions: epiretinal prosthesis and subretinal prosthesis. Each prosthesis technique has its advantages and disadvantages. For epiretinal prosthesis, it is easier to be implanted and has the advantage of keeping most of the electronics in the vitreous cavity, off the retinal surface, which greatly helps in dissipating the heat generated by the implant device. In this paper, a brief overview of retinal prostheses concepts is introduced. After that, several important aspects of epiretinal electrical stimulation will be discussed. Moreover, some practical epiretinal prosthesis devices developed by researchers in United States, Germany and Japan in the past have been reviewed. We hope that the devices will be used widely in the near future.