Objective To investigate the effects of stellate ganglion block(SGB) on the pulmonary artery endothelial nitric oxide synthase (eNOS) and mean pulmonary artery pressure ( MPAP) in rabbits with hypoxic pulmonary hypertension ( HPH) .Methods Twenty-four rabbits of both sexes weighing 2.5-3.0 kg were anesthetized with 1.5% sodium pentobarbital 30 mg? kg-1 . Spontaneous breathing was maintained. Left stellate ganglion was exposed aseptically. An epidural catheter was fixed with one end placed close to stellate ganglion and the other end outside the neck through a hole in the skin for administration of drugs. One week later the rabbits were randomly divided into 4 groups with 6 animals in each group: Ⅰ control group; Ⅱ SGB group; Ⅲ hypoxia group and Ⅳ SGB + hypoxia. In group Ⅲ and Ⅳ the animals were placed in a closed box filled with hypoxic air (O2 % = 10 ? 2% ) 8 h a day for 2 weeks. In group Ⅱ and Ⅳ 0.25% bupivacaine 0.5 ml was injected through the catheter 3 times a day for 3 days. In group Ⅰ and Ⅲ normal saline 0.5 ml was injected instead of bupivacaine. The effect of SGB was confirmed by ptosis and miosis. The content of eNOS in pulmonary artery was detected using immunohistocheistry technique. Pulmonary artery was cannulated after thoracotomy for determination of MPAP. Results There was no significant difference in MPAP between control and SGB groups. MPAP was significantly increased in hypoxia and hypoxia + SGB groups compared with control group(P

Objective To evaluate the safety and feasibility of injection test which is used to locate esophageal balloon catheter.Methods A prospective study was conducted. The patients undergoing invasive mechanical ventilation (MV) admitted to general intensive care unit (ICU) of Beijing Tiantan Hospital Affiliated to Capital Medical University from May 2015 and March 2017 were enrolled. The commercially available esophageal balloon catheter was modified to perform injection test. The catheter was withdrawn step by step and the injection test was repeated until the presence disturbance wave presented, which indicated that the balloon had just entered the esophagus. The position where disturbance wave appears was named 0 cm. End-expiratory occlusions were performed at the positions of+15,+10,+5, 0, -5, -10 and -15 cm, respectively, and the changes of esophageal pressure (Pes) and airway pressures (Paw) were measured in the spontaneous breathing and passive ventilation, and the ratio between the changes (ΔPes/ΔPaw) was calculated.Results A total of 20 patients were enrolled, of which 15 patients finished both the spontaneous and the passive ventilation parts, and 2 patients finished only the spontaneous part and 3 patients finished only passive part. ① Disturbance waves could be induced by injection test in all patients. The average depth of disturbance wave in spontaneous breathing was deeper than that in passive ventilation (cm: 42.4±3.8 vs. 41.8±3.3), but there was no significant difference between the two ventilation settings (P = 0.132). No adverse events occurred during the study period. ② Pes increased with the stepwise withdraw of esophageal catheter, reached the maximal value at+5 cm, and then decreased when the catheter was further withdrawn, no matter in the spontaneous or the passive ventilation. In spontaneous breathing, the ΔPes/ΔPaw was within the ideal range (0.8-1.2) at the positions of 0, -5 and -10 cm. The ΔPes/ΔPaw was closest to unity at the positions of 0 cm (0.98±0.15). The ΔPes/ΔPaw at -15 cm (0.66±0.26) was significantly lower than that at 0 cm (P < 0.05). For passive ventilation, the ΔPes/ΔPaw was within the ideal range at the positions of -5 cm and -10 cm, and the ΔPes/ΔPaw was closest to unity at the positions of-10 cm (0.94±0.12). The ΔPes/ΔPaw at 0 cm and -5 cm was significantly higher than that at -10 cm (1.43±0.31 and 1.12±0.14, respectively); while the ΔPes/ΔPaw at -15 cm (0.68±0.23) was significantly lower than that at -10 cm (allP < 0.01).Conclusions Ideal position of the esophageal balloon catheter could be determined quickly and easily by using injection test. The method is safe and clinically feasible.Clinical Trial Registration Clinical Trials, NCT02446938.

The Caco-2 cell model was widely applied in the research of absorption,metabolism and toxicity of drugs, especially in the aspect of anticancer,inorganic and traditional Chinese medicine,It has become an important tool of the study on medicine.

Objective To evaluate the effect of heme oxygenase-1 (HO-1) protein transduction on acute lung injury in septic rats.Methods Eighteen healthy male Sprague-Dawley rats,aged 7-9 weeks,weighing 210-260 g,were randomly allocated into 3 groups (n =6 each) using a random number table:sham operation group (group Sham),sepsis group (group Sep),and fusion protein PEP-1-HO-1 group (group HO).Sepsis was produced by cecal ligation and puncture (CLP).In group HO,PEP-1-HO-1 fusion protein 0.6 mg was injected via the left iliac vein at 1 h before CLP and 5 h after CLP.The equal volume of normal saline was given instead of PEP-1-HO-1 in Sham and Sep groups.At 12 h after CLP,blood samples were collected from the right common carotid artery for measurement of serum tumor necrosis factoralpha (TNF-α) and interleukin-6 (IL-6) concentrations.The rats were then sacrificed,and lungs were removed for microscopic examination and for determination of wet/dry lung weight ratio (W/D ratio) and malondialdehyde (MDA) content (by thiobarbituric acid colorimetric method).Results Compared with group Sham,the W/D ratio and MDA content were significantly increased,the serum TNF-α and IL-6 concentrations were significantly increased (P＜0.05),and the pathological changes were significantly aggravated in Sep and HO groups.Compared with group Sep,the W/D ratio and MDA content were significantly decreased,the serum TNF-α and IL-6 concentrations were significantly decreased (P＜0.05),and the pathological changes were significantly attenuated in group HO.Conclusion HO-1 protein transduction can attenuate acute lung injury in septic rats,and the mechanism is probably related to inhibition of lipid peroxidation in lung tissues and systemic inflammatory responses.

Objective To compare the similarities and differences of the five detection methods used in the detection of fungi in vaginal secretions,and find the most sensitive、the most specific、the fastest、the most cost effective and the simplest method used in the detection of fungi in vaginal secretions.Methods A total of 442 patients were selected from the Department of Gynecology of Shenzhen OCT Hospital from May 2016 to August 2016.The vaginal secretion of 442 specimens was detected by using the methods of fungi culture、saline and KOH suspension method,Gram stain,Wright''s stain and Vaginitis Multi Test Kit.In these five methods,Fungi culture were using as gold standard to evaluate the specificity,sensitivity,negative predictive value,positive predictive value and accuracy of the other four methods.Results Using the fungus culture method to detect 442 cases of vaginal secretion,we found the positive rate of mycotic infection was 34.8%(154/442).Compared with the fungi culture method,the Specificity of saline and KOH suspension method was 97.9%,the sensitivity was 64.9%,the negative predictive value was 83.9%,the positive predictive value was 94.3% and the accuracy was 86.4%;the Specificity of Gram stain was 96.5%,the Sensitivity was 83.1%,the negative predictive value was 91.4%,the positive predictive value was 92.7% and the accuracy was 91.8%;the Specificity of Vaginitis Multi Test Kit was 84.7%,the Sensitivity was 46.8%,the negative predictive value was 74.8%,the positive predictive value was 62.0% and the accuracy was 71.5%;the Specificity of Wright''s stain was 96.9%,the Sensitivity was 78.6%,the negative predictive value was 89.4%,the positive predictive value was 93.1% and the accuracy was 90.5%.Conclusion Gram stain could be the most sensitive and specific method in the four methods,with highest accuracy,and the the fastest,the most cost effective and the simplest method for the detection of fungi in vaginal secretions.The accuracy of detecting fungi in vaginal secretions could be improved by the combination of Gram stain method in clinical work.

Objective To investigate the effects of galantamine on the myocardial ischemia-reperfusion (I/R) injury in rats and the possible mechanism.Methods Fifty male SD rats weighing 225-275 g were randomly assigned into 5 groups (n =10 each):sham operation group (group SH); I/R group; galantamine + I/R group (group GAL); M receptor antagonist atropine + galantamine + I/R group (group AT); vagus nerve cut-off + galantamine + I/R group (group VGT).Myocardial I/R was induced by occlusion of left anterior descending branch of coronary artery for 30 min followed by 120 min reperfusion.Normal saline 2 ml/kg was slowly injected via the femoral vein at 30 min before ischemia in groups SH and I/R.Galantamine 4 mg/kg was slowly injected via the femoral vein at 30 min before ischemia in group GAL.Atropine 4 mg/kg was slowly injected via the femoral vein at 45 min before ischemia in group AT and the other procedures were the same as those in group GAL.Bilateral cervical vagus nerves were cut off at 45 min before ischemia in group VGT and the other procedures were the same as those in group GAL.At the end of reperfusion,the hearts were removed for determination of myocardial infarct size,MPO and SOD activities,and MDA contents.Results The myocardial infarct size was significantly larger,the MPO activity and MDA content were significantly higher,and the SOD activity was significantly lower in group I/R than in group SH,and in groups AT and VGT than in group GAL (P ＜ 0.05).The myocardial infarct size was significantly smaller,the MDA content and MPO activity were significantly lower,and the SOD activity was significantly higher in group GAL than in group I/R P ＜ 0.05).Conclusion Galantamine has protective effect on myocardium against I/R injury and regulation of peripheral vagus nerve tension may be involved in the mechanism.

BACKGROUND: Natural bilogical bone-derived materials processed with physical and chemical methods possess natural network pore system. They have good cellular compatibility, and help osteoblasts attach and grow on them, and can be used as the scaffolds of osteoblasts.OBJECTIVE: To observe the osteogenetic effect of partially deproteinised decalcified bone (PDDB) as scaffolds of osteoblasts in vivo.DESIGN: A randomized and controlled trial.SETTING: Central Laboratory of the First Hospital of Kunming Medical College.MATERIALS: Partially deproteinised decalcified bone; human embryonic periosteum-derived osteoblasts; twenty 4-week-old BALB/c nude mice,with the body mass of 25 to 28 g, of either gender.METHODS: This experiment was conducted at the Central Laboratory of the First Affiliated Hospital of Kuming Medical College from January to June 2003. PDDB composited with human embryonic periostea-induced osteoblasts were implanted into the nude mice after cultured for 1 week in vivo, 4 scaffolds in each nude mouse. Composite of scalfolds and cells implanted on the left side of the spine was set as experimental side and simple implanted material on the right side of the spine was set as blank control side. Then alkaline phosphatase activity and routine histological examination were performed 4 and 8 weeks after operation.MAIN OUTCOME MEASURES: General observation, alkaline phosphatase activity and routine histological examination were performed after the materials were taken out.RESULTS: Twenty nude mice entered the stage of result analysis. ① General observation of the implanted materials: No necrosis, ecpyesis, or fluidifying was found around the implanted materials, but ingrowth and enwrapption of soft tissues were found. ② Measurement of alkaline phosphatase activity: alkaline phosphatase activity after PDDB composited osteoblasts in vivo was stronger at week 8 than at week 4 [(22.854±6.018) nkat/g vs(11.286±4.268) nkat/g], and was much stronger than that of the simple implanted materials [(1.217±0.083) nkat/g vs (2.717±0.583) nkat/g]. ③ Results of routine histological examination: Cartilage formed at week 4 and part of cartilage formed new bone and marrow cavity at week 8 at the experimental side, cartilage and new bone formed much more as time went by, but there was no any cartilage or bone formation at the control side.CONCLUSION: Cartilage and bone form after PDDB composited with osteoblasts are implanted, and more cartilage and new bone form as time passes. PDDB can be used as the scaffolds of osteoblasts.

Objective To investigate the effect of PEP-1-heme oxygenase-1 (PEP-1-HO-1) fusion protein transduction on hypoxia-reoxygenation (H/R) injury in rat H9c2 cells. Methods After construction of the prokaryotic expression plasmid pET15b-PEP-1-hHO-1 containing the human heme oxygenase-1 gene, it was then transformed to make PEP-1-HO-1 fusion protein express. The H9c2 cells were cultured in high-glucose Dulbecco's modified Eagle's medium (DMEM) supplemented with 15% fetal bovine serum and randomly divided into 4 groups (n = 4 each): control group (group C), H/R group, low-concentration fusion protein group (group L-HO), and high-concentration fusion protein group (group H-HO). The cells were exposed to 22 h of hypoxia followed by 8 h of reoxygenation. PEP-1-HO-1 fusion protein was added to the culture medium with a final concentration of 1.0 μ mol/L (group L-HO) or 2.0 μmol/L (group H-HO) before hypoxia. The cells and supernatant of the culture medium were collected after reoxygenation to determine the activity of lactate dehydrogenase (LDH) in the supernatant and the content of malondialdehyde (MDA) and activity of superoxide dismutase (SOD) in the cells. Results The SOD activity was significantly lower, while the MDA content and LDH activity were significantly higher in group H/R, L-HO and H-HO than in group C (P ＜0.05). The SOD activity was significantly higher, while MDA content and LDH activity were significantly lower in group L-HO and H-HO than in group H/R, and in group H-HO than in group L-HO ( P ＜ 0.05). Conclusion PEP-1-HO-1 fusion protein transdution can protect H9c2 cells against H/R injury in rats.

Objective To evaluate the effects of heme oxygenase-1 (HO-1) mediated by cell penetrating peptide PEP-1 on liverinjury induced by intestinal ischemia/reperfusion (I/R) in rats.Methods Twenty-four male Sprague-Dawley rats,aged 7-9 weeks,weighing 210-260 g,were randomly divided into 3 groups (n =8 each):sham operation group (group S),intestinal I/R group (group I/R) and PEP-1/HO-1 group (group HO).To establish a model of intestinal I/R,intestines were exteriorized and the superior mesenteric artery was exposed and occluded for 45 min ischemia,and then the clamp was removed for 120 min reperfusion.The PEP-1/HO-1 fusion protein 0.5 mg was injectedvia ihe left iliac vein 30 min prior to ischemia in group HO.The superior mesenteric artery was only exposed but not occluded in group S.At the end of reperfusion,blood samples were collected from the right common carotid artery for measurement of serum aspartate aminotransferase (AST),alanine aminotransferase (ALT) activities.The rats were then sacrificed and livers were removed for microscopic examination and for determination of malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in livertissues.Results Compared with group S,serum AST and ALT activities and MDA content in liver tissues were significantly increased,while SOD activity in liver tissues was decreased in groups I/R and HO (P ＜ 0.05).Compared with group I/R,serum AST and ALT activities and MDA content in liver tissues were significantly decreased,while SOD activity in liver tissues was increased in group HO (P ＜0.05).Liver injury induced by intestinal I/R was significantly attenuated in group HO compared with group I/R (P ＜ 0.05).Conciusioon HO-1 protein mediated by cell penetrating peptide PEP-1 can attenuate liver injury induced by intestinalI/R in rats.

Objective To investigate the effects of heme oxygenase-1 (HO-1) transduced by cell penetrating peptide PEP-1 on renal ischemia/reperfusion (I/R) injury in rats.Methods Eighteen male Sprague-Dawley rats,aged 7-9 weeks,weighing 210-260 g,were randomly divided into 3 groups (n =6 each):sham operation group (group S),renal I/R injury group (group I/R) and fusion protein PEP-1/HO-I + I/R group (group HO).I/R injury was produced by occluding bilateral renal arteries for 45 min followed by reperfusion for 6 h.The fusion protein PEP-1/HO-1 was injected via the left iliac vein 30 min prior to ischemia in group HO.Bilateral renal arteries were only exposed but not occluded in group C.Blood samples were collected from the right common carotid artery at 6 h of reperfusion for determination of serum blood urea nitrogen (BUN) and creatinine (Cr) concentrations.The malondialdehyde (MDA) content,superoxide dismutase (SOD) activity and HO-1 expression in renal tissues were measured.Results Compared with group S,the levels of MDA,serum BUN and Cr were significantly increased,the SOD activity was decreased,and HO-1 expression was up-regulated in groups I/R and HO (P ＜0.05).Compared with group I/R,the levels of MDA,serum BUN and Cr were significantly decreased,the SOD activity was increased,and HO-1 expression was up-regulated in group HO (P ＜ 0.05).Conclusion HO-1 protein can be successfully transduced into renal tissues by PEP-1 and transduced HO-1 protein reduces renal I/R injury by inhibiting lipid peroxidation response.