Objective To Observe the adjusting effect of bone morphogenetic protein 4 (BMP4) on cholinergic-expression in embryo rats' neurons. Method Neurons isolated from hippocampus and cerebral cortex of 15-20 days gestational age SD rat's brain were cultivated in a DMEM/F12 medium containing Ara-C and identified with morphological characters and microtubulin associated protein 2 (MAP2 ) immunocytochemical test. The medium was replaced with BMP4 96 hours after Ara-C adding, and cultivated another 10 days. The expression of choline acetyltransferase (ChAT) of the adjusted cells was observed by immunocytochemical staining and FITC-labeled cholinergic nerve cells Flow Cytometry. Results It has been showed that cultured neurons accorded with the typical morphological characters and the immunologic markers of neurons; the percentage of ChAT-positive cells in BMP4 group was prominently higher than that in control group(13.3?1.37% vs 6.44?0.81%, P

AIM To invesigate the effect of subarachnoid transplantation of APA microcapsulized bovine chromaffin cells (BCCs) on mRNA expression for Nav1.8 in the dorsal root ganglia neurons(DRG) of rats with neuropathic pain by means of in situ hybridization. METHODS SD rats were randomly divided into four groups of five. Normal rats were used as control group (group C). Rats with right sciatic nerve been ligated were used as CCI group. Five to six hundred empty APA microcapsules(group APA) or 5?10 6 APA microcapsulized BCCs (group APA-BCCs) were grated into subarachnoid space of CCI rats 7 days after operation. Allodynia and hyperalgesia were measured by Von-Frey filaments and CO 2 laser 7 days after transplantation. DRG in lumbar four and five was taken out and 15 ?m freezing sections were made 7 days after tansplantation. Sections was used to detect mRNA expression for TTX-resistent Na + Nav1.8 by in situ hybridization with Dig-labeled RNA probe. RESULTS The mRNA hybridization signal for Nav1.8 in DRG of group CCI and group APA was lower than that of group C. The expression of mRNA for Nav1.8 in DRG was higher in group APA-BCCs than that in group CCI and group APA with abatement of allodynia and hyperalgesia. There was no difference in the mRNA hybridization signal for Nav1.8 in DRG between group APA-BCCs and group C. CONCLUSION mRNA expression for Nav1.8 in DRG of CCI ratswas down-regulated. APA microcapsulized BCCs grafting can reverse the down-regulation of mRNA expression for Nav1.8 in DRG of CCI rats. Restoration of mRNA expression for Nav1.8 in DRG contributes to the analgesic effect of subarachnoid transplantation of APA microcapsulized BCCs.

Objective To construct a cell line with long time secreting endostatin. Methods Human endostatin cDNA containing interleukin-2 (IL-2) secreting peptide was cloned into eukaryotic expression plasmid pSNA2.0 to construct recombinant plasmid pSNA2.0/Endostatin. The plasmid pSNA2.0/Endostatin was transfected into HEK293 cells by cationic liposome. The positive cell clones were selected by G418 and named hED/293. The expression of endostatin protein was analyzed by Western-blot. Results Endostatin could be determined in the supernatant of hED/293 cells. Conclusion The recombinant eukaryotic expression vector is correctly constructed. The human endostatin protein can be expressed and secreted.

Objective To establish the model of glioma by hypodermic of C6 cells line into the nude mice and observe the growth characteristics. Methods The C6 gliocyte suspension was injected into the left subaxillary hypoderma of nude mice. The diameter and the volume of the tumor were measured and calculated, and the volume-time growth curve was plotted. The cells of tumor were observed under the HE and glial fibrillary acidic protein (GFAP) immono-histochemistry staining. Results The tumor appeared in inoculation area 7 d after injection, and presented nearly exponential development after about 20 d. The survival time of the nude mice is 22～48 d. The tumors were with sharp border, affluent blood vessel, and a great number of GFAP-positive cells. Conclusion The model of glioma can be well induced by hypodermic of the cultured C6 gliocyte into nude mouse.

BACKGROUND: Heme oxygenase-1 (HO-1) promotor region has a pair of dinucleotide(guanosine thymidine, GT) repeats with a lengthy polymorphism, also named microsatellite polymorphism. Experiments in vitro have shown that we can indirectly learn about the level of gene transcription by measuring the number of GT repeats.OBJECTIVE: To investigate if an association exists between restenosis after percutaneous coronary intervention(PCI) and microsatellite polymorphism in HO-1 gene promoter.DESIGN: A case-control study based on the observation of the patients with coronary heart disease after undergoing coronary stenting.SETTING: Wards of the department of cardiology of a university hospital.PARTICIPANTS: A total of 118 patients were admitted from April 1996 to May 2002 at the Department of Cardiology of the First Hospital of Peking University who underwent successful coronary stenting. Inclusion criteria: The patients with coronary heart disease who underwent coronary stent implantation for more than 3 months now came to perform coronary angiography in follow-up. Exclusion criteria: Angiography showed that the stenosis of lumen in diameter in the patients with coronary heart disease was less than 50%and the follow-up in angiography was less than three months. There were 92males and 26 females aged(62±10) years old and the informed consents were obtained. The patients were divided into two groups according to the criteria stipulated by American Heart,Lung and Blood Association: in-stent restenosis(68 cases) and non-restenosis (50 cases).METHODS: DNA of the peripheral blood was isolated from the whole blood. The length of GT repeat was confirmed by PCR amplification and Spreadex Gel electrophoresis. Selected samples were sequenced with Sanger's method.MAIN OUTCOME MEASURES: Microsatellite gene frequency of HO-1promoter and its relationship with restenosis RESULTS: Patients with GT repeats <25 GT in the HO-1 gene promoter on either allele had significantly less often restenosis than patients without (47.5% vs. 68.4% ,P<0.05). After controlling some possible confound ing factorsfor coronary heart diseases, multivariate analysis indicated that still there was a significant difference between the two groups in restenosis rate(odd ratio 0. 418,95% CI: 0. 197 to 0. 887,P<0.05).CONCLUSION: The present study indicated that short(GT) n repeats of HO-1 gene promoter is associated with reduced post-PCI restenosis, which suggests the genetic contribution to in-stent restenosis after stent implantation. It may have important meanings to prevent the occurrence of restenosis.

AIM: To investigate the effects of non-ventilated lung with N_2O on systemic oxygenation and lactic acid level in arterial blood during one lung anesthesia. METHODS: Twenty-two patients, ASA Ⅰ-Ⅲ, scheduled for selective pulmonary surgery, were randomly divided into two groups: control group (group A, n=11) and observation group (group B, n=11). Group A: the non-ventilated lung was kept open to the air; group B: N_2O 2 cmH_2O through CPAP system was insufflated into the non-ventilated lung during one lung ventilation. The anesthesia was induced with intravenous midazolam (0.05 mg?kg~(-1)), propofol (0.5-1.0 mg?kg~(-1)), fentanyl (4 ?g?kg~(-1)), and vecuronium (0.1 mg?kg~(-1)) and was maintained with inhaling isoflurane. Blood gas analysis and lactic acid was recorded 20 min after two-lung ventilation (TLV) in the supine position, 20 min after one-lung ventilation (OLV) in the supine position, 20 min and 40 min after OLV in the lateral position and at the end of operation and the shunt fraction was calculated. RESULTS: PaO_2 in group B was significantly higher than that in group A (P

Objective To investigate the biomechanicai characteristics of flexor profundus tendons repaired after decimeter wave therapy, and to observe the effect of decimeter wave therapy on early active mobilization. Methods A total of 56 Leghorn chickens were randomly divided into a therapy group and a control group with 28 chickens in each. The 3rd and 4th toes of their left feet were employed for the establishment of a tendon injury model. The flexor profundus tendons were cut and repaired. Gypsum support was applied and fixed with an adhesive plaster after the operation. The operated sites on toes Ⅲ and Ⅳ were exposed. The external fixation was removed 3 weeks later and the chickens were left free to move. Decimeter wave therapy ( frequency 915 MHz, power 8 Watts) was ap-plied for 10 minutes once daily on the left foot of each chicken in the therapy group from day 1 until 3 weeks after the operation. Sham decimeter wave therapy was applied to chickens in the control group. Four chickens from each group were randomly selected at the 1st, 7th, 10th, 14th, 18th, 21st and 28th days for biomechanical analysis. Biome-chanical parameters including tensile strength of rupture (Pmax), elongation ratio at rupture (δimax) and the tensile adhesion strength of the rupture zone (W0>) were observed at each time point. Results At the 7th, 10th, 14th, 18th, 21st and 28th day after the operation, the differences in Pmax, δmax and W0 between the therapy and control groups were statistically significant. The results of the therapy group were better than those of the control group. Conclusions Local decimeter wave therapy after flexor tendon repair can promote intrinsic healing and reduce ex-trinsic healing. The speed and quality of healing are improved. The elasticity and tenacity of the injured tendons are enhanced. Therefore decimeter wave therapy is helpful for early active mobilization training.

Objective To observe the effect of APA-BCC(alginate-polylysine-alginate microencapsulated bovine adrenal medullary chromaffin cells)microcapsules transplantation into the subarachnoid space of cancer pain patients on endogenous opioid peptides and catecholamine concentration in cerebrospinal fluid(CSF).Methods The different doses of APA-BCC microcapsules were transplanted into the spinal subarachnoid space of cancer patients with moderate or serious pain.The concentrations of Leu-enkephalin(L-EK),β-endorphin(β-EP),dynorphin A(DynA),noradrenaline(NA)and adrenaline(AD)in CSF were tested.Results L-EK concentration in CSF increased remarkably after transplantation,and the increase was most remarkable when transplantation doses were 1.0×107and 1.25×107;there were no remarkable changes of β-EP,DynA,NA and AD after transplantation.Conclusion The analgetic effects of APA-BCC microcapsules tranplantation may correlate with the increase of L-EK in CSF of cancer pain patients;the dose of 1.0×107 and 1.25×107cells may be the most effective dose.

BACKGROUNDRRM1 may be a prognostic factor in non-small cell lung cancer (NSCLC). The aim of this study is to evaluate RRM1 expression and prognosis in NSCLC by the means of tissue microarray.

METHODSA total of 417 paraffin-embedded specimens of NSCLC from Lung Cancer Study Center in Guangdong Provincial People's Hospital were collected and tissue microarray was constructed. RRM1 expression was detected by SP method and its correlation with prognosis was evaluated.

RESULTSNo statistic difference was found in RRM1 expression in different gender, age, tumor site, histology, differentiation, T stage, N stage, M stage and pTNM stage groups (P > 0.05). Univariate analysis showed that RRM1 was not an independent prognostic factor (P > 0.05). At the multivariate analysis, differentiation and N stage were considered independent prognostic factors.

CONCLUSIONSRRM1 expression detected by immunohistology is not an independent prognostic factor in NSCLC. TNM stage is still the best prognostic factor up to now.

Objective To investigate the application value of limited liquid resuscitation combined with continuous renal replacement therapy (CRRT) in treatment of severe acute pancreatitis (SAP) companied with abdominal compartment syndrome (ACS).Methods The retrospective cohort study was adopted.The clinical data of 67 patients with SAP companied with ACS who were admitted to the Fuling Center Hospital of Chongqing from January 2005 to December 2014 were collected.Among 67 patients,33 receiving conventional liquid resuscitation between January 2005 and December 2010 were allocated into the control group and 34 receiving limited liquid resuscitation combined with CRRT between January 2011 and December 2014 were allocated into the observation group.Observation indicators included:(1) required fluid volume and time of negative fluid balance in the 2 groups;(2) changes of pathological and physiological indicators after treatment in the 2 groups;(3)outcomes and prognosis of patients in the 2 groups.Measurement data with normal distribution were represented as (x)±s and comparison between groups was analyzed using t test.Count data were analyzed using the chi-square test.Repeated measurement data were analyzed by repeated measures ANOVA.Results (1) Required fluid volume and time of negative fluid balance in the 2 groups:required fluid volumes at 6,24,48 and 72 hours after resuscitation were (2 449±339) mL,(4 820±757) mL,(9 428± 1 272) mL,(13 127± 1 565) mL in the control group and (2 360± 314) mL,(4 582±530) mL,(8 564± 970) mL,(11 470± 1 253) mL in the observation group,respectively,with a statistically significant difference in required fluid volume between the 2 groups (F=13.035,P＜0.05) and in required fluid volume at 48 and 72 hours between the 2 groups (t=3.132,4.794,P＜0.05).Time of negative fluid balance in the observation group and control group was (4.3± 1.7)days and (6.4 ±1.8)days,respectively,showing a statistically significant difference between the 2 groups (t =4.913,P＜0.05).(2) Changes of pathological and physiological indicators in the 2 groups after treatment:time factors:from 0 h to 168 h postoperatively,APACHE Ⅱ score,C-reaction protein (CRP),D-dimer,IAP,Bla and oxygenation index were changed from 20.9±4.1 to 13.9±2.6,from (167±39)mg/L to (55±17) mg/L,from (1 652±1 544) μg/L to (993±500)μg/L,from (23.4±3.4)cmH2O (1 cmH2O=0.098 kPa) to (21.4±2.9)cmH2O,from (4.6±1.6) mmol/L to (1.4±0.5)mmol/L,from (189±27) mmHg (1 mmHg =0.133 kPa) to (152±23)mmHg in the control group,and chaged from 21.5±5.1 to 11.0±2.8,from (168±36)mg/L to (44±19)mg/L,from (1 634±1 525) μg/L to (578±350) μg/L,from (23.2±2.5)cmH2O to (17.4±2.6)cmH2O,from (4.5±1.6)mmol/L to (0.8±0.3)mmol/L,from (188±26)mmHg to (196±25)mmHg in the observation group,respectively,showing gradual decreasing with time and statistically significant differences between the 2 groups (F =186.415,581.118,34.618,212.416,262.272,207.645,P＜0.05).Treatment factors:there were no significant differences in changing trends of APACHE Ⅱ score,D-dimer and Bla between the 2 groups (F=3.499,2.350,3.516,P＞0.05),and there were significant differences in changing trends of CRP,IAP and oxygenation index between the 2 groups (F=4.009,15.276,14.959,P＜0.05).Interaction effect between time factors and treatment factors:there were obviously interaction effects between time factors and treatment factors in APACHE Ⅱ score,CRP,IAP and oxygenation index (F=4.890,4.912,23.874,28.615,P＜0.05) and no interaction effects between time factors and treatment factors in D-dimer and Bla (F=2.803,1.920,P＞0.05).(3) Outcomes and prognosis of patients in the 2 groups:numbers of patients with surgery,local complications and infection and duration of hospital stay were 11,16,14,(46±17)days in the control group and 4,6,6,(36±14) days in the observation group,respectively,with statistically significant differences between the 2 groups (x2=4.484,7.221,4.910,t =2.433,P＜0.05).Mortality and hospital expenses were 24.2％ (8/33),(33± 18) x 104 yuan in the control group and 8.8％ (3/34),(27± 14)× 104 yuan in the observation group,respectively,with no statistically significant difference between the 2 groups (x2 =2.901,t =1.283,P＞ 0.05).Conclusion Limited liquid resuscitation combined with CRRT can effectively control IAP of patients with SAP companied with ACS and improve oxygenation index,meanwhile,it can also reduce number of patients with surgery,infection and local complications and duration of hospital stay.