Cardiovascular Diseases ; diagnosis ; genetics ; Humans ; MicroRNAs ; blood ; genetics

Objective Endogenous cadiac stem cells become the famous star in the cardiac regeneration field in recent years.But the biological behavior of cardiac stem cells has not been fully explained clearly.This experiment intends to research the cardiac stem cell niche and its characteristics,as well as the succinct method for cardiac stem cells (Cardiospheres derived cells) culture in vitro.Methods Hearts from four 8-week-old SD rats were cut into pieces less than I mm3.Then,the small tissues were digested by trypsogen and collagenase in turn.And the undigested myocardium was collected and cultured in media until the de novo cells bespread the pertri-dish bottom.The 3rd generation of cardiac stem cells were harvested and prepared for immunofluorescence to detect the expression of related molecular markers,so as to identify the differentiation of these stem cells.Then,the cultural tissues were collected and embedded in paraffin,and the H-E staining,Masson staining,immunoflnorescence and TUNEL apoptosis assay were carried out.In addition,EdU was added into the medium and cuhured for 72 hours before the cultural tissues were harvested so as to label the proliferative cells.Results After 7 ～ 10 days in culture,some small,round and phase-bright cells aresed from the adherent plated tissue.These cells had the spontaneous differentiation potency in vitro culture.The immunofluorescence shows mitotic phase cells were c-kit positive,and most of passage cells were α-sarcomeric actin 、cardiac troponin T、flk-1 and vemintin positive while CD90 negative,but few of them were α-smooth muscle actin 、conexin-43 、CD31 and CD45 positive.There were lots of newborn cells grow exuberantly while accompanied apoptosis of myocardial cells in the paraffin section.The myocardial collagen remodels in the cultural myocardial tissues at the same time.The proliferative cells in the cultural tissues were EdU positive.Throe myocardium within the cultural tissues were MMP-2、MMP-9 and TGF-β1 positive,On the contrary,the proliferative cells were almost negative when stained with those antibodies.Conclusion Conclusions It's a convenient way to harvest cardiac stem cells by the use of myocarlial? tissues cultured in vitro.Those stem cells grow and proliferate in the cultural myocardial tissues accompanied with the degradation of extracellular matrix.Cardiospheres derived stem cells have the spontaneous differentiation potency when cultured in vito,and should be a promising? seed cells for stem cell therapy.

Objective Both underuse and overuse of anticoagulant therapy may lead to severe adverse effects. Emerging portable monitering devices, which provides reliable and accurate measurements, were reported to be potential alternatives to tra-ditional monitering recta-analysis regimens. This study was intended to evaluate the effects of serf-monitoring or serf-management (self-testing and serf-dosing) of anticoagulant as compared with that of traditional monitoring. Methods Relevant trials reported before October 2008 were identified in a number of electronic database and analyzed with software RevMan 4.2. The primary out-comes included death from any cause, major bleeding event, thromboembolic event and the proportion of patients whose interna-tional normalized ratio (INR) were within the therapeutic range. Results Seventeen RCT of serf-monitoring were identified.Pooled estimates revealed significant reductions in the thromboembolic events (odds ratio 0.46, 95% CI0.33 -0.64), all-cause mortality (0.61,0.40 -0.93), and major haemorrhage (0.80, 0.58 - 1.10) with self-monitoring as comparing with traditional monitoring. No difference was noted in minor haemorrhage. 15 trials reported improvements in the mean proportion of patients whose INR were within target range. Conclusion Self-management regimen is superior to traditional monitoring in the outcomes of oral anticoagulation. Patients capable of self-monitoring and serf-adjusting have fewer thromboembolic events and lower mortali-ty than those undergoing self-monitoring alone. However, self-monitoring requires education and training for patients.

Objective To study the changes of gastrointestinal movement function in rats with chronic unpredictable mild stress(CUMS) and explore the mechanisms underlying it.Methods The rats were divided into stress model group and control group.The stress model rats were induced by 21-day chronic unpredictable mild stress as well as social-isolated fed.The rate of ink propulsion of gastrointestinal tract and the contraction of intestinal canal in rats were observed.Immunohistochemistry was adopted to detect the expression of OT in rats.Results (1) After the models were induced,weight-gain and sucrose preference of model group ((69.97 ± 9.81) g,(49.05± 5.98) g) were lower than those in control group ((116.27 ± 13.60) g,(83.51 ± 3.08) g) (P ＜ 0.001),and both the crossing-score and rearing-score ((24.00 ± 13.52),(3.90 ± 2.51)) were lower than those in control group ((53.60 ± 27.98),(11.50 ± 8.85)) in the open-field test.(2) The rate of ink propulsion of model group ((67.33 ± 6.24) ％) was decreased when compared to the control group ((76.83 ± 10.00) ％) (P ＜ 0.05),and the intestinal canal contraction amplitude and contraction frequency ((1.37 ± 0.18) g,(0.58 ± 0.02) S-1) were lower than those in control group ((1.88 ± 0.13) g),(0.62 ± 0.04) S-1) (P ＜ 0.05).(3) Compared with the control group (6.07 ± 3.71),OT immunoreactive substance was increased in model group (59.17 ± 16.08) of rats (P＜0.001).Conclusion Chronic stress can cause the decrease in gastrointestinal movement function of rats.These changes may be related to the increased expression of OT in paraventricular nucleus.

OBJECTIVETo summarize the clinical effect of avulsed skin replantation of hand and foot via vacuum sealing drainage (VSD) combing low temperature technique.

METHODSFrom March 2012 to October 2013,13 cases with avulsed skin replantation of hand foot using combined technique included 8 males and 5 females with an average age of 32 years old ranging from 18 to 62 years. The time from injury to hospital was 1 to 4 hours (2.4 hour in average). The reasons of injury included machine injury in 7 cases and rolling over by cars in 6 cases. The parts of injuried involved finger in 2 cases,back of the hand in 5 cases and dorsum of foot in 6 cases. The area of avulsed skin was 5 cm x 6 cm to 12 cm x 16 cm,tendon and bone exposure was found in 4 cases. VSD was operated in all patients and the avulsed skin was refrigerated in the temperature of -4 °C or -80 °C. After 4 days, the skin stored in the -4 °C was replanted to the wounded place in 5 cases and in 3 cases the skin was planted to the donor site of flap. The skin stored in the -80 °C was replanted in 4 cases after 7 or 8 days, 1 case after 45 days.

RESULTSOf the 13 cases, 1 case of degloved injury from lower leg to dorsal foot,the replanted skin was necrosis completely; 1 case of degloving injury with fourth finger,the skin which replanted after 45 days survived approximately 30%,cured after skin-graft many times. In the other cases, the survival area of replanted skin was more than 85%, all cured after dressing. According to the standard of skin survival area evaluation by Jia et al, 11 cases showed excellent, 1 showed medium and 1 showed inferior. There were no complication about grafted skin rupture after the skin survived in 11 patients,after 4 to 22 months follow-up, the resiliency of grafted skin showed good. Sensation recovery was measured by BMRC standard: 3 cases of S3, 5 cases of S3, 3 cases of S2.

CONCLUSIONVSD combining lower temperature technique in skin replantation provides time and space for wound preparation and treatment plan for the patients who need second surgery, especially for the large area skin degloving,this method could utilize the degloved skin efficiently, decrease the donor site area, alleviate the pain and financial burden,reduce the scar formation of donor site and impediment.

Adolescent ; Adult ; Cryopreservation ; methods ; Drainage ; instrumentation ; methods ; Female ; Foot Injuries ; surgery ; Hand Injuries ; surgery ; Humans ; Male ; Middle Aged ; Replantation ; Skin ; injuries ; Skin Transplantation ; Young Adult

Ectopic thyroid gland is a kind of congential deformity which occurs during thyroid embryo development. Combined with this case, our study introduced the diagnosis, treatment process and the postoperative outcome of "intracardiac ectopic thyroid" in the case, discussed the causes of this kind of disease from the perspective of tissue and embryo, and summarized the basic characteristics of this disease. Through the research and analysis of our study, we come to the conclusion and wish to offer some suggestions: although this disease is relatively rare, it should also be paid attention to to avoid misdiagnosis and mistreatment if encountered middle-age female patients with the intracardiac mass located in the right ventricle outflow tract.

OBJECTIVETo study DNA damage, Bcl-2 and Bax expression, and ultrastructure change in spermatogenic cell of mice by cadmium exposure.

METHODSTwenty-four male mice were divided into 4 groups: 3 groups treated with cadmium chloride of 1, 5, 10 micromol x kg(-1) x d(-1) i.p. respectively for 5 days, and one normal saline control group. The DNA damage of spermatogenic cell by single-cell gel electrophoresis technology was detected. The expression positive rate of Bcl-2, Bax protein in spermatogenic cell by the immunohistochemical method was assayed, and the ultrastructural change of spermatogenic cell by the transmission electron microscope was observed.

RESULTSDNA damage rates of of spermatogenic cell in 1, 5, 10 micromol/kg cadmium chloride groups were higher than that of normal group (P < 0.001). Bcl-2 protein expression positive rates were lower than that of normal group (P < 0.001). Bax protein positive expression rate in 5 micromol/kg group was higher than those in normal group, and 1, 10 micromol/kg groups. The ultrastructure of karyotis, karyotheca, mitochondria, endoplasmic reticulum in three treated groups had different degree of damage and the degree of ultrastructural change was increasing with rising concentration of cadmium.

CONCLUSIONCadmium exposure will cause the DNA break, Bcl-2 and Bax protein abnormal expression and ultrastructural change in spermatogenic cell.

Animals ; Apoptosis ; Cadmium Chloride ; toxicity ; DNA Damage ; Male ; Mice ; Mice, Inbred ICR ; Proto-Oncogene Proteins ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; Spermatozoa ; drug effects ; metabolism ; ultrastructure ; bcl-2-Associated X Protein ; metabolism

Objective　To investigate the effect of fatty acid binding protein 7 （FABP7） on blood-brain barrier （BBB） permeability in cerebral ischemia/reperfusion （I/R） rats and the mechanisms involved. Methods　BBB cell model was established by co-culture of rat brain microvascular endothelial cells and astrocytes. The BBB model of I/R injury was established by using oxygen-glucose deprivation and reoxygenation （OGD/R） method. This model was treated with the FABP7 recombinant protein （rh-FABP7） alone or together with S7155.ENDOHM instrument，Western blotting and flow cytometry were used to determine the transendothelial electrical resistance （TEER），FABP7，TJ and matrix metalloproteinase （MMP） protein levels，and apoptosis. The rat brain I/R model was established，and rh-FABP7 was injected intraperitoneally. The neurological function score，wet and dry weight method，and Evans blue staining were used to determine the neurological function，brain water content，and BBB permeability of rats. Results　FABP7 overexpression reversed the OGD/R-induced down-regulation of TEER，and FABP7 and TJ protein expression，as well as the up-regulation of MMP2/9 expression and apoptosis （P<0.05）. S7155 treatment promoted the effect of FABP7 on MMP2/9 and TJ protein levels，TEER，and apoptosis （P<0.05）. Moreover，FABP7 overexpression reversed the I/R-induced increase of neural function score，brain water content，and BBB permeability and decrease of FABP7 protein level in rats（P<0.05）. Conclusion　FABP7 alleviated the damage of brain I/R to BBB integrity by inhibiting MMP2/9.

Objective To develop a HPLC method for determining 14 hydrolyzed amino acids in Pfaf fia .Methods The sample was derivatized with phenyl isothiocyanate (PITC) .Amino acids were separated on Waters XBridge Shield RP18 (4.6 mm × 250 mm ,5 μm) column at the flow rate of 0.8 ml/min ,detected at 254 nm .The column temperature was 25 ℃ . Results The response was linear for 14 amino acids with a correlation coefficient r>0 .9990 .The average recoveries (n=6) were 90 .2%-105 .1% .Amino acids derivative solution remained stable in 24 hours .Conclusion This well-established method is very reliable .It can be used as a quantitative determination method for 14 amino acids in Pfaf fia .

Complex systems exist widely,including medicines from natural products,functional foods,and bio-logical samples.The biological activity of complex systems is often the result of the synergistic effect of multiple components.In the quality evaluation of complex samples,multicomponent quantitative analysis(MCQA)is usually needed.To overcome the difficulty in obtaining standard products,scholars have proposed achieving MCQA through the"single standard to determine multiple components(SSDMC)"approach.This method has been used in the determination of multicomponent content in natural source drugs and the analysis of impurities in chemical drugs and has been included in the Chinese Pharmacopoeia.Depending on a convenient(ultra)high-performance liquid chromatography method,how can the repeatability and robustness of the MCQA method be improved?How can the chromatography conditions be optimized to improve the number of quantitative components?How can computer software technology be introduced to improve the efficiency of multicomponent analysis(MCA)?These are the key problems that remain to be solved in practical MCQA.First,this review article summarizes the calculation methods of relative correction factors in the SSDMC approach in the past five years,as well as the method robustness and accuracy evaluation.Second,it also summarizes methods to improve peak capacity and quantitative accuracy in MCA,including column selection and two-dimensional chromatographic analysis technology.Finally,computer software technologies for predict-ing chromatographic conditions and analytical parameters are introduced,which provides an idea for intelligent method development in MCA.This paper aims to provide methodological ideas for the improvement of complex system analysis,especially MCQA.