Objective To investigate the effects of substance P on anoxia/reoxygenation (A/R) injury to cardiomyocytes of neonatal rats.Methods Cardiomyocytes of neonatal rats were isolated from Sprague-Dawley rats,aged 1-3 days,and were cultured in 6-well plates for 72 h.The cardiomyocytes were then assigned into 4 groups (n =3 each) using a random number table:control group (group C),A/R group,substance P group (group SP) and substance P + D-SP group (group SP + D-SP).The cells were cultured routinely for 6 h in group C and the cells were exposed to 99.9 ％ N2 in an incubator at 37 ℃ for 3 h followed by reoxygenation for 2 h in the other groups.The cells were incubated with substance P with the final concentration of 10-7 mol/L for 1 h before anoxia in group SP.The cells were incubated for 1 h with substance P with the final concentration of 10-7 mol/L and D-SP (a specific antagonist of neurokinin-1 receptor) with the final concentration of 10-6 mol/L before anoxia in group SP + D-SP.The apoptosis rate and lactate dehydrogenase (LDH) activity were detected at the end of reoxygenation using TUNEL assay and LDH assay kit,respectively.Results Compared with C group,the apoptosis rate and LDH activity were significantly increased in A/R,SP and SP+ D-SP groups.Compared with A/R group,the apoptosis rate and LDH activity were significantly decreased in SP and SP + D-SP groups.Compared with SP group,the apoptosis rate and LDH activity were significantly increased in SP + D-SP group.Conclusion Substance P can attenuate A/R injury to cardiomyocytes of neonatal rats,and activation of neurokinin 1 receptors is involved in the mechanism.

Objective To evaluate the effect of calcitonin gene-related peptide (CGRP) on anoxiareoxygenation (A/R)-induced injury to neonatal rat cardiomyocytes incubated in high glucose medium.Methods Cardiomyocytes were obtained from 1-3-day old Sprague-Dawley rats and cultured in the culture medium containing 15％ bovine calf serum and then seeded onto 6-well plates at a density of 10 × 105/ml (3 ml/well).The cells were randomly divided into 5 groups (n =9 each):normal glucose medium control group (NG group),high glucose medium group (HG group),high glucose medium + A/R group (HG+ A/R group),high glucose medium + A/R + CGRP group (HG + A/R + CGRP group),and high glucose medium + A/R + CGRP+ CGRP8-37 group (HG + A/R + CGRP + CGRP8-37 group).The cells were incubated in normal glucose (5.5 mmol/L) medium for 72 h in NG group.In HG group,the cells were incubated in high glucose (25.0 mmol/L) medium for 72 h.In HG + A/ R group,the cells were incubated in high glucose medium for 72 h and then exposed to 3 h of anoxia followed by 2 h of reoxygenation.In group HG + A/R + CGRP,the cells were incubated in high glucose medium for 72 h,CGRP (final concentration 10-8 mol/L) was then added to the culture media and the cells were incubated for 1 h and then underwent A/R.In HG + A/R + CGRP + CGRP8-37 group,the cells were incubated in high glucose medium for 72 h,CGRP (final concentration 10 8 mol/L) was then added to the culture media and the cells were incubated for 1 h and then underwent A/R.In HG + A/R + CGRP + CGRP8-37 group,the cells were incubated in high glucose medium for 72 h,CGRP8-37 (final concentration 10-8 mol/L) and CGRP8-37 (CGRP receptor antagonist,final concentration 10-7 mol/L) was then added to the culture media and the cells were incubated for 1 h and then underwent A/R.Apoptosis in cardiomyocytes was detected using TUNEL and apoptosis index (AI) was calculated.The lactate dehydrogenase (LDH) activity in the culture medium was analyzed.Results AI and LDH activity were significantly higher in HG group than in NG group,and in HG + A/R group than in HG group.Compared with HG + A/R group,AI and LDH activity were significantly decreased in HG + A/R + CGRP group,while no significant changes were found in HG + A/R + CGRP + CGRP8-37 group.Compared with HG + A/R + CGRP group,AI and LDH activity were significantly increased in HG + A/R + CGRP + CGRP8-37 group.Conclusion CGRP attenuates A/R-induced injury to neonatal rat cardiomyocytes incubated in high glucose medium via combing with CGRP receptor.

Objective To observe the change of cerebral oxygen metablism and the level of blood lactate in early stage of sepsis in rats.Method Sixty-four SD rats were randomly(random number)divided into septic group and control group.The sepsis model of rat was made by lipopolysaccharide (LPS,10 mg/kg)injected intra-abdominally,and rats of control group were treated with the same amount of physiological saline instead.And each group was further divided into 4 sub-groups of4 h,6 h,12 h and 24 h after treatment.At each interval,blood samples were obtained via jugular vein for detecting blood oxygen saturation (Sjv02)and blood lactate(LA).The blood oxygen saturation(Sa02)of ventral aorta was also measured.Arteriovenous oxygen content difference (AVD02) and oxygen extraction fraction (OEF) were studied.These four variables were analyzed and compared between two groups.Results The AVD02 and OEF in sepsis group were higher than those in control groups of 3 h,6 h and 12 h (P<0.05) ,but lower than those in control group of24 hours (P<0.05).The Sjv02 in sepsis group was lower than that in control groups of 3 h,6 h and 12 h (P<0.05),but increased to that in control group of 24 h (P>0.05).LA in sepsis group was higher than that in control group in each interval (P<0.05).Conclusions In early stage of sepsis,cerebral perfusion,cerebral oxygen supply and cerebral oxygen extraction increase.The global oxygen metabolic variables(Sjv02,AVD02 and OEF)and the level of LA can be used to objectively and accurately evaluate the cerebral oxygen metabolic dysfunction in early stage of sepsis.

Objective To evaluate the changes of respiratory mechanics and central drive in sleeping patients with chronic obstructive pulmonary disease (COPD).Methods A total of 14 patients with moderate to severe COPD (age 63.7± 6.4 years) and 10 healthy volunteers,admitted to our hospital from October 2010 to October 2011,were chosen in our study.After performing routine pulmonary function test,all subjects were monitored by respiratory mechanics,diaphragm electromyogram (EMGdi) and polysomnography (PSG) in waking and sleeping state.The different physiological parameters were recorded and calculated on a continuous basis in time sequence for 30 minutes.Results In sleeping state as compared with waking state,there were no differences(P＞0.05) in respiratory rate(RR) and the ratio of inspiratory time to duration of one breath (Ti/Ttot),but there were reductions (P ＜ 0.05) in the tidal volume (VT),minute ventilation (VE),mean inspiratory volume per second(VT/Ti),dynamic lung compliance(CLdyn),root mean square (RMS)and pulse oxygen saturation(SpO2).Meanwhile there were significant increases (P＜0.05) in airway resistance(Raw) and pressure-time product(PTP) in COPD group.In normal group,there were no differences (P＞0.05) in all above physiological parameters in sleeping state compared with waking state.Conclusions There are abnormal changes in respiratory mechanics characterized by increased airway resistance and work of breathing,meanwhile in reduced central drive and requirement of ventilation,which may be the main reasons for hypopnea and hypoxemia in sleeping COPD patients.

Objective To study the clinical value of anti-ribosomal P0 protein antibodies (anti-P0). Methods Line immuno-assay and immunoblotting methods were used to detect anti-P0 and rRNP antibodies of 49 SEE patients and 61 patients with other rheumatic diseases.The expressions of anti-P0 were compared with those of rRNP antibodies.Possible relations between anti-P0 and clinical features and other antibodies in SLE were explored.Results 36.7%and 6.1% of SEE patients showed positive anti-P0 and rRNP antibodies respectively.Anti-P0 was always negative in patients with other rheumatic diseases (P＜0.01).Among SLE pa tients,incidence of skin rash was 77.8%(35.5%) ff anti-P0 was found to be positive (negative) (P＜0.05). Besides,for patients with positive and negative anti-P0,the chance of positive anti-SmD1 was 61.1% and 19.4% respectively (P＜0.01).Sensitiviy of anti-P0 for SLE diagnosis was 36.73% with specificity and positive predictive value as high as 100%,and 66.30% as the negative predictive value.Conclusion Sera anti-P0 is highly specific for SLE and it is associated with the occurence of rash and the expression of anti-StuD 1 anti bodies in SEE.

Objective To study the effect of-1 site single nucleotide polymorphism(SNP) on CCNH gene promoter transcription activity.Methods PCR and site-directed mutagenesis technology were used to construct CCNH basic promoter and-1G mutate promoter.Dual-Luciferase Reporter assay system was used to detect the transcription activity of constructed promoter.Results In AD293 cells,the activity of-1G mutate type promoter was significantly lower than that of wild type-1T promoter(P

Two patients underwent liver transplantation of blood type incompatibility were collected from Fuzhou General Hospital of Nanjing Military Area Command of Chinese PLA. Case 1: A male who had primary hepatic carcinoma underwent classic orthotopic liver transplantation; the blood of donor was type A, and the blood of recipient was type O. Case 2: A female having history of type B hepatitis underwent classic orthotopic liver transplantation due to pregnancy combining with severe liver disease and coagulation disorder; the blood of donor was type B, and the blood of recipient was type O. Immunohistochemistry staining was used to observe pathological changes and deposition of various immunoglobulin and complement in two cases following liver transplantation of blood type incompatibility under optic microscope and to investigate diagnostic standard of humoral rejection. The results showed that linear or granular depositions of IgG, IgM, IgA, C4c, C4d, and Clq were found in endothelial cells of hepatic sinusoid, suggesting that IgG and other immunoglobulin exhibited a strongly positively diffused deposition on the endothelial cells of hepatic sinusoid, while expression of C4d and other complements was also found. All those mentioned above could be considered as evidences to prove onset of humoral rejection in transplanted liver tissue.

Objective To investigate the possible existence of HBoV in children with acute respiratory infections in Nanjing area and explore its relationship with clinical characteristics.Methods A total of 397 nasopharyngeal secretion samples were collected from children with acute respiratory infection,admitted from July 2009 to June 2010 in Nanjing Children'S Hospital affiliated to Nanjing Medical University,and 50 cases of children without symptoms of respiratory infection were recruited as control group,whose nasopharyngeal secretion samples were also collected.HBoV was determined by real-time fluorescence quantitative PCR.MP and CT were detected by real-time fluorescence quantitative PCR in those HBoV-positive samples.RSV,ADV,IVA,IVB,PIV-1,PIV-2,PIV-3 and hMPV were detected by direct antigen-specific immunofluorescence assays.HBoV NP-1 fragments were amplified and sequenced in 5 HBoV positive samples randomly selected.The results were compared with the known GenBank sequence,and thereby the phylogenetic tree was established.The epidemiological characteristics,clinical presentation and the final clinical diagnosis of HBoV were analyzed according to the clinical data of the HBoV-positive patients.Results Thirty-three HBoV-positive cases were detected by real-time fluorescence quantitative PCR method with a positivity rate of 8. 3% ( 33/397 ). Among the 33 HBoV-positive cases, 19 cases (57.6%) were multiple infections with HBoV and other pathogens, the top three of which were MP (27.3% ,9/33 ),RSV (24.2% , 8/33 ) and PIV-3 ( 12. 1% ,4/33 ). Affected children aged from 7 to 36 months old accounted for 75.8% of the total ( 25/33 ). The measured HBoV NP-1 gene sequences of 5 specimens were consistent,indicating a high homology (99% to 100% ) with the stl, st2 and WHL-1. Conclusions HBoV is one of the pathogens of children's acute respiratory infections in Nanjing. HBoV NP-1 gene is highly conserved,with little variation in different seasons and in different regions and therefore can be used as a marker for real-time fluorescence quantitative PCR and other methods.

Objective To evaluate the value of the clinical application of piezosurgery-assisted sliding genioplasty.Methods A total of 9 patients (7 women,2 man,aged 19-40 years) with microgenia were performed operation by intraoral V incision with subperiosteal exposure of the mandibular mentum area.Note relaxtion and preservation of the mental nerves.A horizontal osteotomy of cortical bone on anterior mandible was underwent by the conventional electric bone saw and internal plate bone was sectioned with the piezoelectric device.The segment was divided with preservation of the lingual soft tissue attachment and was advanced in the mandibular mentum area.These was fixed in the reposition by titanium plate and screw for preventing displacement.Results The piezoelectric device produced selective sectioning of the mineralized bone structures,bone was cut precisely without damage to vital anatomic structures and dental pulp,and was much easier to control and less aggressive,creating only minimal damage to soft tissues.The operative effects were satisfactory during 3- 6 months of follow-up.Stable chin contours were created and became nature.Iatrogenic problems with bleeding,in fection,mental nerve injury had been avoided.Conclusions Piezosurgery is a safe and precise technique to reduce the complication rate and to improve the treatment outcomes of genioplasties.

Objective To investigate a safe and effective method of lengthening the short nose.Methods Thirty-nine cases of short nose were corrected by the bilateral septal spreader graft and columellar strut using autologous cartilage,combined with expanded polytetrafluoroethylene (e-PTFE)to reconstruct the supporting structures to improve the forward and upward strength of the nasal tip in order to increase the nasal length and to correct the over rotation of nasal tip.The shield and cap grafts were also used for the patients whose nasal tip were too low with vertical dome division technique.Results All the patients showed preserved nasal length after surgery with well-proportioned nasal features and the nasal lip angles were normal.There was no evidence of postoperative nasal shortening.No operative or postoperative complications were observed by follow-up for 6 months to 1 year,except for 2 cases with graft cartilage being apparent on nasal tip.All the patients were satisfied with the surgical results achieved.Conclusions It is necessary to provide powerful forward and upward strength to lengthen nose and correct the over rotation of nasal tip effectively and safely.The application of bilateral septal spreader graft and columellar strut using autologous cartilage and combined with e-PTFE represents a valuable tool for the short nose and proper cartilage grafts of nasal tip can improve the height of the nasal tip and further correct the over rotation of the nasal tip.