Objective To explore the effect of hypoxia on cultured rat Sertoli cells in vitro.Methods In a Sertoli-germ cells co-culture system,the effect of hypoxia on the morphological characteristics of Sertoli cells were observed,including the number of Sertoli cells,the cytoskeleton changes after Coomassie brilliant blue staining and the detachment of germ cells.Results The loosing and retracting of cytoskeleton in Sertoli cells was found.Both survival rate of Sertoli cells and detachment of germ cells showed time-response relation.Conclusion Hypoxia damages Sertoli cells.

This is a prospective study on early discovery and diagnosis to cerebral palsy(CP).Fivehundred and forty cases with high risk neonates who were born from July 1. 1993 to December 31. 1994 infour districts in Beijing were monitored for one year.Eight cases were diagnosed for CP from 38 suspiciouscases by examining specific primitive reflex. nerve reflex.muscular tension. early symptoms and CT orMRI.etc. The study indicates that it is very important for decreasing CP incidence to strengthen healthmeasures in perinatal period and to avoid premature infant birth.

Objective To investigate cell proliferation and invasiveness of cervical cancer HeLa229 cell after knockdown of NF-?B signaling pathway by P65 siRNA.Methods RNA interference was employed for specific inhibition of the expression of P65.HeLa229 cell was divided into transfected group and untransfected group.Cell viability was detected by MTT after the HeLa229 cells were transfected with or without P65 siRNA for 24,48,72 h.The sensitivity to 5-Fu of the HeLa229 cell,transfected with or without P65 siRNA,was evaluated also by MTT.Boyden chamber experiment in vitro was used to detect the invasion of HeLa229 cell.Results P65 siRNA inhibited the cell proliferation as compared with the untransfected cells.Proliferations of both cells transfected with and without P65 siRNA were inhibited in a concentration-dependent manner,while at the same concentration of 5-Fu the viability of transfected HeLa229 cells was significantly suppressed(P

AIM: To explore the mechanism of apoptosis in pulmonary arterial vascular smooth muscle cells (PASMCs) induced by nitric oxide (NO). METHODS: Wistar rat PASMCs were isolated from explants from the intrapulmonary, incubated with NO donor, sodium nitroprussid (SNP) under 12 or 24 hour normoxic and hypoxic conditions. The cell cycle progression and sub-G 1 of PASMC were analyzed via flow cytometric staining of propidium iodide, and the expression of nuclear transcription factor NF-?B and pro-apoptotic factors caspase-3 were detected using immunochemistry staining. Meanwhile gel electrophoresis of extracted DNA by PASMC was performed. RESULTS: SNP induced PASMC apoptosis in a dose-dependent manner ( P

Objective To explore the effect of the cancer pain nursing team in standardized nursing inter-vention of pain in elderly cancer patients.Methods 76 patients in Department of Pain in our hospital were selected in our research.A random number table method was used to divide them into the control group and the observation group,with 38 cases in each group.Conventional treatment and nursing care was given to control group.The observa-tion group received routine treatment and nursing care by cancer pain nursing team,which the content of nursing was same,but the nursing method is different.Before and after the intervention,impact on pain intensity,adverse reactions and satisfaction for medical and nursing were compared between the two groups.Results Before nursing intervention, the pain score between the two groups had no statistically significant difference(P >0.05).After nursing interven-tion,pain score of the observation group was 1.5 (1,2)points,which was lower than 2(1,3)points in the control group,the observation group had better analgesic effect than the control group (Z = -4.385,P =0.000).The incidence rate of adverse reaction of the observation group was 13.16%,which was lower than 47.37% of the control group,the difference between the two groups was statistically significant(χ2 =10.537,P =0.001).The satisfaction for medical nursing process in the observation group was (9.74 ±0.45 )points,which was higher than (9.42 ± 0.76)points of the control group (t =2.213,P =0.030).Conclusion For senile cancer patients,the cancer pain nursing team could implement the standardized nursing intervention to relieve the body pain level,reduce the adverse drug reactions,so as to improve the quality of life.

Objective To explore the method of isolation, identification and in vitro differentiation of islet stem cells in neonatal rat. Methods The whole pancreata of neonatal rats were digested with collagenase, then,under the (condition) of pH 7.4～7.6,the digested tissue fragments were cultured with serum and serum-free RPMI 1640 (( bFGF), EGF, N2).The whole formation process of new islet cell-like cell masses was examined.The insulin (release) test was used to detect islet function. The expression of nestin was tested by immunocytochemistry. Results The nestin positive cells can be found within 36 h of culture of the digested pancreatic fragments.After addition of bFGF,ECF,N2,nestin positive cells proliferated fast and formed new islets-like cell masses after 18～24 d, and (insulin)ssion could be confirmed. Conclusions The nestin positive cells of pancreatic cells possess the character of islet stem cells, and can form islet like cell masses through culture in vitro.

Objective To explore the isolation, culture of nestin positive cells of the pancreas in newborn rats. Methods The whole pancreas of neonatal rats were digested with collagenase, followed by incubation under pH 7. 6 serum RPMI 1640 for 24 - 36 h and then under pH 7. 4 serum free RPMI 1640 (bFGF.EGF 20 ng/ml.1% N2) for 18-24 d. The expression of insulin.glucagon and nestin were detected by immunofluorescence technique. Nestin.CK19 were detected by RT-PCR. Resides Cells attached to the surface of plates after 24 h incubation under pH 7. 6 condition, and islet-like masses were obtained after 18 -24 d incubation. A monolayer of cells grew out after 24 h of passage of islet-like masses. Nestin positive cells was detected after 24 - 36 h incubation, with no expression of insulin and glucagon. Positive cells of insulin and glucagon were detected in islet-like masses after 24 h passage. Nestin positive cells were detected by RT-PCR in islet-like masses after 24 h passage, but no CK19. Conclusion Insulin and glucagon were expressed in islet-like masses after passage. Nestin positive cells in the pancreas of neonatal rats possessed character of islet stem cells.

AIM: To construct signal peptide-canstatin expression vector pEGFP-C1-SP-Can and express secretable mouse canstatin fusion protein in Eca-109 cells.METHODS: Site-directed mutagenesis was used in amplifying the signal peptide of murine plasminogen to construct the plasmid pEGFP-C1-SP.The cDNA of mouse canstatin,obtained from a cloning vector pMD18T-Can by PCR,was inserted into pEGFP-C1-SP to construct a secretable expression vector pEGFP-C1-SP-Can.Constructed plasmid pEGFP-C1-SP-Can was transiently transfected into Eca-109 cells via lipofectamine,and subsequently its secretable expression in the medium of cultured Eca-109 was observed by Western blotting.RESULTS: DNA sequencing and restriction enzyme analysis attested the validity of the constructed plasmids pEGFP-C1-SP and pEGFP-C1-SP-Can.EGFPcanstatin fusion protein was proved to be secretably expressed in Eca-109 by Western blotting.CONCLUSION: It is concluded that the constructed vector pEGFP-C1-SP-Can is valid and capable of expression in Eca-109,these findings provide a basis for testing the function of mouse canstatin and its application in gene therapy.

Objective To evaluate clinical use of tigecycline in hospital patients. Methods Basic diseases, pathologic examinations, concurrent medication, therapeutic efficacy and side effects of 40 patients in Lishui Central Hospital of Zhejiang Province from January 2012 to December 2014 were analyzed retrospectively. Results The effective rate of patients using tigecycline for anti-infection treatment in hospital was 42. 5%. The rates of rational use, basically rational use and irrational use were 17. 5%, 77. 5% and 5. 0%, respectively. Adverse drug reactions occurred in 6 cases of tigecycline use (15. 0%). Conclusion Clinical use of tigecycline in inpatients was basically reasonable in this hospital. The clinical curative effect of tigecycline was good in a variety of infections caused by sensitive bacteria. However, the incidence of adverse drug reactions was high. Attentions should be paid in clinical application.

Objective: To observe the effects of Naomaitong, a compound traditional Chinese herbal medicine, combined with mobilization of bone marrow mesenchymal stem cells (BMSCs) on neuron apoptosis in rats with cerebral ischemia, and to explore the possible mechanism by detecting the expressions of Fas, FasL and caspase-3 proteins. Methods: Two hundred and two SD rats were divided into sham-operated group, untreated group, recombinant granulocyte colony-stimulating factor (rG-CSF) group, Naomaitong group and Naomaitong plus rG-CSF group (combination group). Focal cerebral ischemia was induced by intraluminal middle cerebral artery occlusion using a nylon thread with some modification. Rats in the rG-CSF group and the untreated group were administered with rG-CSF 10 mug/(kg.d) by subcutaneous injection 3 d before and 2 d after the operation respectively, once a day, and rats in the Naomaitong group and the combination group were intragastrically administered Naomaitong before and after the operation until sacrificed. Two, three, seven and fourteen days after operation, count of CD34-positive cells in peripheral blood and CD34 expression in brain tissue were determined. General neural function score (GNFS) was evaluated. Neuron apoptosis, expressions of Fas, FasL and caspase-3 in rat's brain were all measured. Results: Count of CD34-positive cells in peripheral blood and CD34 expression in brain tissue were high in the untreated group, and reached the peak at 3 d and 7 d respectively. CD34 expression in brain tissue was increased in each treated group, especially in the combination group. GNFS was increased at 3 d and 7 d in the untreated group, 7 d and 14 d in the rG-CSF group and the combination group. Expressions of Fas, FasL and caspase-3 were increased 2, 3 and 7 d after operation, while expression of FasL at 2 d in the rG-CSF group, expressions of Fas, FasL and caspase-3 in the combination group were decreased. Expressions of Fas, FasL and caspase-3 at 7 d and 14 d in the combination group were lower than those in the rG-CSF group. Meanwhile, expressions of Fas, FasL and caspase-3 were decreased in each group at 14 d as compared with those at 3 d. Conclusion: There exists interaction between Naomaitong and BMSC mobilization in the effect of improving nerve function and inhibiting neuron apoptosis in rats after cerebral ischemia. It is implied that Naomaitong combined with BMSC mobilization down-regulates the expressions of Fas and FasL in early phase and then inhibits the apoptosis cascade reaction caused by caspase-3, which causes further inhibition of Fas and FasL expression after cerebral ischemia.