Objective To observe the effect of transforming growth factor beta 1 (TGF-β1) on the expression of collagen in rat atrial and ventricular fibroblasts, and to investigate its specific molecular mechanisms. Methods Tissue explant attachment was used to culture fibroblasts obtained from the atrium and ventricle of rat heart, and they were identified with SABC immunocytochemical staining, and then the following experiments were carried out. (1) Hydroxyproline digestion was performed to study the effects of TGF-β1, within different concentrations (0, 5, 10ng/ml) and different action time (6, 12, 24, 48h) on the content of hydroxyproline in rat's atrial and ventricular fibroblasts. (2) Rat's atrial and ventricular fibroblasts were stimulated with TGF-β1 in optimal concentration and action time, the expression of α-smooth muscle actin (α-SMA) was determined with Western blotting, and the expressions of typeand III collagen mRNA were evaluated with reverse-transcription PCR. The contents of hydroxyproline in the respective cells were measured with hydroxyproline determination. Western blotting was used to measure the protein expression of Smad2/3, p-Smad2/3 and Smad7. Results (1) TGF-β1 was shown to stimulate the collagen synthesis in rat's atrial and ventricular fibroblasts, and the optimal stimulus was TGF-β1 concentration 5ng/ml with action time of 24h. (2) After being stimulated by optimal stimulation effect of TGF-β1, the expression of typeand III collagen and p-Smad2/3 increased, while that of Smad7 decreased significantly only in atrial fibroblasts (P<0.05), but not in ventricular fibroblasts. No statistical difference was found in the expression of Smad2/3 between the atrial and ventricular fibroblasts after being stimulated by TGF-β1 under optimal stimulating conditions. Conclusion TGF-β1 can induce dysbolism of collagen of cardiac fibroblasts with abnormal expression of cytoskeletal protein, which may occur more obviously in rat's atrial fibroblasts than in ventricular fibroblasts, and its mechanism may be related with TGF-β1/SMAD pathway.

The study was aimed to establish a standard procedure for human umbilical cord blood bank. The hematopoietic nucleated cells in cord blood were processed by using sedimentation and centrifugation method. After finishing CD34(+) cell counting, hematopoietic progenitor cell assay, microbial culture, infectious disease test and HLA typing, cord blood units were stored in the liquid nitrogen for further application. The results showed that nucleated cells of cord blood were (10.94 +/- 2.74) x 10(8) per unit; recovery rate of nucleated cells was (79.82 +/- 17.76)%. CD34(+) cells in cord blood were counted as (51.62 +/- 30.53) x 10(5) per unit. Eight units of cord blood were thawed after two years of cryopreservation, the recovery rate of nucleated cells, CD34(+) cells and CFU-GM were (91.4 +/- 6.0)%, (84.6 +/- 20.0)% and (85.8 +/- 14.9)% respectively. It is suggested that the methods and procedure reported for processing and cryopreservation of hematopoietic stem/progenitor cells in the human umbilical cord blood is effective.
Antigens, CD34 ; blood ; Blood Preservation ; methods ; Cell Separation ; methods ; Colony-Forming Units Assay ; Cryopreservation ; methods ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; immunology ; Humans

To study the effects of short-term cryopreservation of cord blood hematopoietic cells in liquid nitrogen, the viability and function of cord blood hematopoietic cells were investigated by using each of 8 samples cryopreserved for six months, one and two years after thawing respectively. Nucleated cells (NC) were detected by blood cell analyzer. CD34+ cells were analyzed by flow cytometry, CFU-GM were cultured and detected in vitro, the survival rate was determined by trypan blue staining. The results showed that the differences of recovery rate of NC, CD34+, CFU-GM were nonsignificant at three different cryopreserved times. In conclusion, the short-term storage in liquid nitrogen showed a good effect on cord blood hematopoietic cell without any significant change of activities and number of the cryopreserved hematopoietic cells.
Cell Survival ; Cryopreservation ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; physiology ; Humans

Objective To investigate the clinical effects of dynamic mild hypothermia on patients with severe brain injury. Methods One hundred and seventy-four patients with severe brain injury, admitted to our hospital from 2003 to 2009, were chosen and randomly assigned to mild hypothermia treatment group, short-term mild hypothermia treatment group and control group; their clinical outcomes and complications were recorded. Results The difference on effective rate of the 3 groups was significant (84.48%, 67.24%, 48.27%, respectively, P<0.05); the mortality in the 3 groups was 8.62%, 15.51%, 36.20%, respectively. No significant difference of complications was showed in the 3 groups (46.55%, 46.55% and 48.27%, respectively, P>0.05). Conclusion Dynamic mild hypothermia therapy can improve the outcomes of severe brain injury according to the patients' condition.

Objective To observe the effect of limited laminectomy combined with foraminal stenosis decompression on preventing C5nerve root palsy and improving neurological function. Methods From March,2014 to May,2016,69 patients with multi-segment cervical spondylotic myelopathy underwent surgical treatment in our hospital were included.Thirty-eight patients(group A)were treated with limited lami-nectomy combined with foraminal stenosis decompression and internal fixation,and 31 patients(group B)under-went routine laminectomy and internal fixation.The postoperative neurological recovery rate,cervical curvature index(CCI)and C5palsy rate were recorded and analyzed. Results No spinal cord and nerve injury occurred during the operation.The width of laminectomy was(16.8±2.1)mm in group A,and was significantly less than(21.7±2.5)mm in group B(t=8.849,P<0.001).There was no significant difference in operation time and intraoperative blood loss between two groups(t<0.439,P>0.05).The Japanese Orthopaedic Association (JOA) score increased continuously after surgery in both groups (F>42.996, P<0.05), and no significant difference was found between them at each time point(t<1.021,P>0.05).The cervical curva-ture index improved after surgery(F>86.379,P<0.05),and no significant difference was found between them at each time point(t<0.943,P>0.05).The spinal cord drift distance was(3.6±0.7)mm in group A,and ws signifi-cantly shorter than(2.5±0.5)mm in group B(t=7.602,P<0.001).There was no significantly difference in the neu-rological recovery rate between two groups(t=0.724,P=0.471).The C5palsy rate was lower in group A(5.2%,2/38)than in group B(22.5%,7/31)(χ2=4.514,P=0.034). Conclusion Limited laminectomy combined with foraminal stenosis decompression could relieve the spinal cord com-pression and restrict the excessive back drift, promote the neurological function recovery and reduce the inci-dence of C5palsy.

OBJECTIVETo investigate the effects of rat tail collagen, hair follicle dermal papilla cells and hair follicle epithelium cells on collagen gel contraction in organotypic culture.

METHODSThe hair follicle organotypic culture was prepared with different concentrations of rat tail collagen, different number of dermal papilla cells and hair follicle epithelium cells in DMEM medium, after cultured for 10 days the diameter of collagen gel was measured.

RESULTThe concentration of rat tail collagen, hair follicle dermal papilla cells and hair follicle epithelium cells significantly influenced on collagen gel contraction in organotypic culture (P<0.01). The contraction of collagen gel was negatively related to the concentration of rat tail collagen, while the concentration of dermal papilla cells and hair follicle epithelium cells was positively related to the contraction of collagen gel.

CONCLUSIONThe key factor influencing collagen gel contraction in organotypic culture is the concentration of rat tail collagen, hair follicle dermal papilla cells and hair follicle epithelium cells.

Animals ; Cell Division ; Cells, Cultured ; Collagen ; physiology ; Gels ; Hair Follicle ; cytology ; Rats

OBJECTIVETo investigate the expression of bFGF, ET-1 and SCF in different passages of cultured dermal papilla cells (DPC), and their possible effect on biological behaviour of DPC.

METHODSThe expression of bFGF, ET-1 and SCF in different passages of cultured DPC was detected by immunocytochemistry and in situ hybridization.

RESULTThe expression of ET-1 and SCF in early passages of cultured DPC was stronger, but became negative in late passages (>6 passages). The stronger the expression of ET-1 and SCF in DPC, the higher ability of DPC to induce hair follicle regeneration.

CONCLUSIONThe expression strength of ET-1 and SCF is related to the ability of DPC inducing hair follicle regeneration.

Endothelin-1 ; analysis ; genetics ; Fibroblast Growth Factor 2 ; analysis ; genetics ; Hair Follicle ; chemistry ; cytology ; physiology ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Stem Cell Factor ; analysis ; genetics

To elucidate further sequence selectivity and nature of the binding of anticancer drugs to DNA, the interaction between anticancer drugs, which are minor groove ligands (distamycin A, DM and netropsin, NP) and intercalator (mitoxantrone, MT), and DNA were studied by electrospray ionization mass spectrometry. The 2 : 1 specific complex of DM and AT-rich DNA were observed principally, while only 1 : 1 specific complex of NP and AT-rich DNA were observed. MT specifically binds to GC-rich DNA. In addition, DM binds to DNA containing 5 A/T bases minor groove almost in a 2 : 1 mode and does not bind to DNA containing 3 A/T bases minor groove. NP binds most strongly to DNA containing 4 A/T bases minor groove. The 1 : 1 specific complex of MT and 6-mer DNA was also observed. The result of competitive binding experiment shows that DM binds more strongly to AT-rich DNA than NP does. These results provide bases for investigating the mechanism of interaction between the drugs and DNA and for improving the structure of target drug.
Antineoplastic Agents ; chemistry ; DNA ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods

Objective To observe the efficacy and safety of applying sirolimus-eluting stents in emergency percutaneous coronary intervention (PCI) for the patients with acute myocardial infarction (AMI).Methods In total,220 patients with AMI were enrolled in this study at Shanghai Tongji Hospital, divided into two groups,one with bare-metal stent and the other with sirolimus-eluting stent.Cardiovascular fatality,major adverse cardiac events (MACE) and target vessel revascularization (TVR) were observed one and six months after PCI in the two groups.Results There was no significant difference in overall fatality and MACE in the 1~(st) or 6~(th) months after PCI between the two groups.Three cardiogenic deaths occurred in bare-metal stent group with a fatality of 2.8 percent,and five deaths in sirolimus-eluting stent group with a fatality of 4.5 percent in six months after PCI.However,rate of restenosis in those with sirolimus-eluting stents was significantly lower than that of bare-metal stents (6.0 percent vs 16.1 percent,P

Adolescent ; Adult ; Athletic Performance ; psychology ; Back ; physiology ; Electromyography ; Exercise ; physiology ; Humans ; Imagery (Psychotherapy) ; Male ; Young Adult