BACKGROUND: L-arginine (L-Arg) is the precursor of endogenous nitric oxide (NO), which joins the pain regulation of peripheral myeloid level and above.OBJECTIVE: To study the role and mechanism of NO in nucleus raphe magnus (NRM) in pain regulation and electroacupuncture analgesia.DESIGN: A random control experiment on rats.SETTING: Physiology Department of Medical College of Xianning College.MATERIALS: The experiment was done during May 2002 to March 2003in the Physiology Department of Medical College of Xianning College. The 63 Wistar rats were randomized into 7 groups with 9 in each. ①. Five groups for the experiment of L-Arg dose-effect relationship: Normal saline group and L-Arg 1, 2, 4, 8 mmol groups. ②. Two groups for the experiment of relationship between L-Arg and electroacupuncture analgesia: Normal saline + electroacupuncture group and L-Arg + electroacupuncture group.METHODS: ①Experiment of L-Arg dose-effect relationship: Normal saline group was injected normal saline. L-Arg 1, 2, 4, 8 mmol groups were injected L-Arg 1, 2, 4, 8 mmol respectively. The volume was all 1.5 μL.Then, the hot water (50±0.5) ℃ was used to stimulate the tail once every 10 minutes to cause tail flick for testing the pain threshold. The observation was carried out continuously for 90 minutes. ②. Experiment of relationship between L-Arg and electroacupuncture analgesia: The two groups were injected normal saline and L-Arg 8 mmol respectively. The volume was all 1.5 μL. Ten minutes later, Zusanli (ST 36) of both posterior legs of rats were punctured with electric stimulation. The frequency was 4-16 Hz, and the intensity in an increasing order of 1, 2, 3V given 10 minutes of each. The electroacupuncture was totally 30 minutes, during which, testing the pain threshold 3 times. After needling, the pain threshold was still tested once every 10 minutes, till 90 minutes after injecting L-Arg.MAIN OUTCOME MEASURES: Pain threshold at different time points.RESULTS: The 63 rats entered the result analysis. ① The pain threshold of L-Arg 1 mmol group at all time points was lower than that of the normal saline group, but without statistical significance (P ＞ 0.05). The pain threshold of L-Arg 2, 4, 8 mmol groups was much lower than that of the normal saline group at all time points (P ＜ 0.001), and the effect was lasting to 80 minutes after injecting L-Arg, and with the increase of L-Arg density, the amplitude of pain threshold lowering was increased. ② The pain threshold of L-Arg + electroacupuncture group was much lower than that of the normal saline + electroacupuncture group at the time points of 20-50 minutes (P ＜ 0.01).CONCLUSION: ① The L-Arg injection to NRM can lower the pain threshold and there is a dose-dependent effect in it. ② The electroacupuncture can raise the pain threshold, and the L-Arg injection can weaken the effect of electroacupuncture analgesia. ③ NO in NRM joins in the pathological and physiological processes of pain and electroacupuncture analgesia. Its increase can lower the pain threshold greatly.

AIM: To investigate the effects of propofol on the expression of tissue-type plasminogen activator (tPA) and matrix metalloproteinase 9 (MMP9) in the hippocampus and the cognitive function in neonatal rats.METHODS: The 7-day-old rats were randomly divided into 3 groups: the rats in control (CON) group were intraperitoneally injected with normal saline for 7 d;the rats in single dose of propofol anesthesia (SP) group were intraperitoneally injected with normal saline for 6 d and with propofol on the 7th day;the rats in repeated dose of propofol anesthesia (RP) group were intraperitoneally injected with propofol for 7 d.Blood glucose and blood gas analysis were tested in 6 rats of each group.The rats were randomly selected from each group to isolate the hippocampal tissues at 2 h, 24 h, 48 h, 72 h and 30 d after the last injection.The spatial learning and memory functions of the other rats aged 25 d were determined by Morris water maze.The morphological changes of the hippocampus were observed by HE staining and Nissl's staining.The expression of tPA and MMP9 at mRNA and protein levels was determined by RT-PCR and Western blot.RESULTS: Compared with group CON, the protein expression of tPA and MMP9 in RP group was significantly decreased at each time point, while no significant decrease was observed in SP group except at the time point of 24 h.Compared with CON group, the mRNA expression of tPA and MMP9 was down-regulated obviously in RP group, which was not significantly down-regulated in SP group.From the 3rd training day of Morris water maze beginning, the escape latency was prolonged, and the space exploration time and the number of crossing the original platform location were reduced in RP group compared with CON group and SP group, while no significant difference was observed between CON group and SP group.Compared with CON group, the number of nerve cells reduced and nerve cells arranged in disorder in the hippocampus in RP group.Moreover, the number of Nissl body decreased significantly and finally developed into neuronal degeneration and necrosis in RP group, and no significant difference between SP group and CON group was observed.CONCLUSION: Repeated dose of propofol anesthesia leads to long-term cognitive dysfunction in neonatal rats, which may be related to the down-regulation of tPA and MMP9 expression and destruction of normal morphology and function of neurons in hippocampus, whereas single dose of propofol anesthesia has no such effects.

Aim To observe the improvement effects of parecoxib on ventricular remodeling after acute myocardial infarction in rats and its influence on PI3K/Akt signaling pathway.Methods Forty male Sprague-Dawley rats were randomly divided into five groups(n=8):sham operation group (group S), ventricular remodeling model group(group R), low dose of parecoxib group(group P1), middle dose of parecoxib group(group P2), and high dose of parecoxib group(group P3).A myocardial infarction model was established by ligating the left anterior descending branch(LAD) of coronary artery in group R, group P1, group P2 and group P3.One day after the operation,the rats were given intraperitoneal injection of 4,8,12 mg·kg-1 parecoxibin Group P1, group P2 and group P3,respectively, for two weeks.The same volume saline was given in group S and group R.Four weeks later, LVSP, LVEDP,+dp/dtmax,-dp/dtmax were monitored.The hearts were harvested to calculate left ventricular hypertrophy index.The pathological change of heart was examined with an optical microscope.The expressions of atrial natriuretic peptide(ANP) mRNA and brain natriuretic peptide(BNP) mRNA were detected by RT-PCR.The expression of PI3K,Akt,P-Akt and caspase-3 was detected by Western blot.Results Compared with group S, the cardiac function was decreased, the left ventricular hypertrophy index, the expression levels of ANP,BNP mRNA, caspase-3 were increased, and the expression levels of PI3K, P-Akt were reduced in group R(all P<0.05).Compared with group R, the cardiac function was ameliorated, the left ventricular hypertrophy index were reduced in group P2 and group P3(all P<0.05).The expression levels of ANP,BNP mRNA, Caspase-3 were decreased, and the expression levels of PI3K and P-Akt were increased in group P1,group P2 and group P3(all P<0.05).Conclusions Middle and high doses of parecoxib can mitigate the process of ventricular remodeling after myocardial infarctionand improve the myocardial function, and its underlying mechanism may be related to activating PI3K/Akt signaling pathway.

Aim To investigate the role of matrix metalloproteinase-9 down-regulation in the learning and memory dysfunction induced by propofol treatment in rats.Methods 7-day-old SD rats were randomly divided into three groups(n=18):control group(NS group) and repeated doses of propofol group(RP group) was intraperitoneally injected with normal saline and propofol respectively for consecutive seven days, single dose of propofol group(SP group) were intraperitoneally injected with normal saline first for consecutive six days, and then injected with propofol on 7th day.The blood gas and glucose levels were monitored of six rats randomly selected from each group.Morris water maze was conducted to test the learning and memory functions of the remaining rats.The expression of MMP-9, BDNF and caspase-3 was detected by Western blot, and the hippocampal neuron apoptosis was determinated by TUNEL staining.Results Compared with NS group and SP group, the escape latency in RP group was prolonged significantly, exploration time and the number of crossing the platform in RP group were markedly decreased(P<0.05).The expressions of MMP-9 and mBDNF in RP group declined, but the expression of proBDNF and the ratio of proBDNF/mBDNF in RP group were higher than those in NS group and SP group(P<0.05).Compared with NS group and SP group, the number of apoptotic neurons and the expression of cleaved-caspase-3 in RP group were increased significantly, but the expression of pro-Caspase3 in RP group was reduced(P<0.05).There was no difference between SP group and NS group regarding all the results(P>0.05).Conclusions Repeated exposure to propofol can lead to a decline in long-term learning and memory functions in neonatal rats, which may be related to the down-regulation of MMP-9 expression, proBDNF and mBDNF conversion disorder in hippocampus and the apoptosis of hippocampal neurons.However, single exposure to propofol has no significant effect.

Objective To assess the value of CT angiography (CTA) in diagnosis of re-entry tears in type B aortic dissection. Methods One hundred and ten patients with typical type B aortic dissection were enrolled. Data derived from the CTA scans of all the patients and DSA of 70 patients were reviewed. The number, location and size of the re-entry tears were calculated and analyzed. CTA findings were compared with DSA results by using Kappa statistics. Results The mean number of re-entry tears was 3.41±1.90, and 83.64% (92/110) patients had multiple tears. Re-entry tears occurred in the middle of the abdominal aorta were more common than that in the other palce of aorta. The large ones commonly located in the most remote position of dissection. The sensitivity and specificity of finding re-entry tears with CTA was 93.78% and 88.89% respectively, the Kappa value was 0.827 (P＜0.01). Conclusion The consistency between CTA and DSA was good. The re-entry tears in type B aortic dissection can be depicted clearly with CTA, and CTA can be used as the main method for further observation and follow-up of this disease.

Objective In this paper,we analyzed the initial review projects of clinical research during past five years in a hospital,which aims to summarize the experience and analysis existent problems,in order to provide a reference for the future hospital ethics review.Methods Summarizing and analyzing the meeting records during past five years,which were also in the field of the initial review projects of clinical research.Results Clinical trials of drug and divice,and clinical research projects exist in varying degrees of problems,which express in these two aspects:research design and ethical rational.Conclusions Ethics Committee should stengthen review the following two parts,the scientific of research project and the informed consent form.Improve their own system constantly,and ensure the efficient operation of the Ethics Committee.

Objective To investigate the effect of 1?,25-dihy-droxyvitamin D3 (1?,25(OH)2D3) on the expression of receptor activator of NF-?B ligand (RANKL),osteoprotegerin (OPG) and RANKL mRNA,OPG mRNA,of rat osteobalsts (OB). Method The expression of RANKL and OPG was detected by the method of Immunohistochemistry and ELISA. RANKL mRNA and OPG mRNA were determined through FQ-PCR. Results Compared with the control group and the group with 1 nmol/L 1?,25(OH)2D3,10 and 100 nmol/L 1?,25(OH)2D3 can significantly induce the expression of RANKL and RANKL mRNA. 1,10 nmol/L 1?,25(OH)2D3 can stimulate the expression of OPG and OPG mRNA significantly,while 100 nmol/L 1?,25(OH)2D3 can inhibit the expression of OPG mRNA significantly. There were no difference in the rate of RANKL/OPG and RANKL mRNA/OPG mRNA between the control group and the group with 1 nmol/L 1?,25(OH)2D3,however the rate of RANKL/OPG and RANKL mRNA/OPG mRNA in the group with 10,100 nmol/L were higher than the control group and the group with 1 nmol/L 1?,25(OH)2D3. Conclusion Lower dosage of 1?,25(OH)2D3 (1 nmol/L) had no significant effect on bone turnover,but higher dosage of 1?,25(OH)2D3 (10,100 nmol/L) can enhance bone turnover through facilitating the formation and activity of osteoclasts via enhancing RANKL mRNA/OPG mRNA and RANKL/OPG..

Objective In order to study the biological characteristics of macrophages and provide the materials to study the survival mechanism of intracellular parasites, we conducted this study to establish a high-purity alveolar macrophage isolation and culture method.Methods Goat lungs were lavaged with normal saline in sterile environment several times, and cells were collected and then goat alveolar macrophages were purified by density gradient centrifugation using peripheral blood mononuclear cells (PBMC) solution.The isolated goat alveolar macrophages were cultured in cell culture medium containing 10% fetal bovine serum and cell morphology was observed under an inverted microscope every day,and the phagocytic activity of the cells was detected by chicken red blood cell phagocytosis test.Flow cytometry was used to detect CD14, a characteristic monocyte-macrophage surface marker.Results The adherent cells were characterized by typical macrophage morphology, pseudopodia and protrusions, showing round and irregular shape, rich cytoplasm, and large cell body.Of the cultured macrophages, 54.5% could phagocytize chicken erythrocytes and showed good phagocytic activity.After one month of in vitro culture, 93.7% of the cells were able to express CD14 antigen, which had a macrophage-specific immunophenotype.Conclusions The alveolar macrophages obtained in this study have high purity and good bioactivity, thus provide a cell model for studying the immune mechanism of intracellular parasites.

Objective To investigate the surgical methods and experience of laparoscopic radical cystectomy and orthotopic ileal neobladder for invasive bladder cancer. Methods The clinical data of 14 patients with invasive bladder cancer underwent laparoscopic radical cystectomy and orthotopic ileal neobladder were collected retrospectively during March 2011 and October 2014. Results The 13 patients with invasive bladder cancer were successfully completed laparoscopic radical cystectomy and orthotopic ileal neobladder. 1 case was treated with laparotomy because of unsatisfactory surgery ifeld caused by excessive tumor bleeding. Twelve cases of the urethra-neobaldder anastomosis were completed through the abdominal incision, while for the other 2 cases, the anastomosis was done under the laparoscope, 2 cases were performed neovesicourethral anastomosis using single-needle running sutures through laparoscopy. The median operative time was 444 minutes, the mean intraoperative blood loss was 490 ml. Postoperative pathologic results conifrmed that 12 cases were bladder transitional cell carcinoma (1 case with partial squamous cell carcinoma) and 2 cases with bladder adenocarcinoma. No severe complication occurred except for 2 cases of urinary leakage and 1 case of urinary incontinence. Patients were followed up for 6-56 months,within which 3 patients were died of distant metastasis, 1 case was detected with intracranial metastasis, 1 case was found with urethra-vesical anastomotic stenosis while cured after urethrotomy. Ten cases were well recovered and the mean volume of the neobladder was 300 ml. Conclusions Laparoscopic radical cystectomy and orthotopic ileal neobladder have the advantage of better therapeutic effects, safety, minimal invasion and rapid recovery, which are the preferred therapeutic methods for invasive bladder cancer.

OBJECTIVE: To explore the high risk factors of adult complex appendicitis, and to provide a reference for the development of a reasonable treatment strategy for acute appendicitis. METHODS: A retrospective case-control study was conducted to collect clinical data of 312 adult patients with acute appendicitis confirmed by pathology undergoing appendectomy, including open and laparoscopic surgery, from May 2011 to August 2016 at Affiliated Hospital of Qingdao University. Age <14 years old, pregnant women, complicating abscess around the appendix, AIDS, blood system diseases, autoimmune diseases, inflammatory bowel disease or progressive cancer patients were excluded. According to the intra-operative findings and pathological types, patients were divided into complex appendicitis(112 cases, including gangrene and perforation) and non-complex appendicitis (200 cases, including simple and non-perforated appendicitis, ie suppurative appendicitis). After comparing the clinical data of these two groups, statistically significant variables were induded for multivariate logistic regression analysis to identify risk factors of complex appendicitis, and to establish a regression model. Enter method was applied to establish the regression equation: P=ExpiΣBiXi/1+ExpΣBiXi, and to calculate the relative risk of each variable. Meanwhile, retrospective and prospective verification was performed on this predictive model (cases of acute appendicitis from September 2016 to December 2017 were further collected). The sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of complex appendicitis were calculated with the regression model. RESULTS: Comparison of the clinical data between the complex appendicitis group and the non-complex appendicitis group showed that differences of 10 preoperative indexes were statistically significant, including period from abdominal pain to admission [(59.1±42.6) hours vs. (47.5±34.4) hours, t=3.051, P=0.002], white blood cell count [(12.9±3.7)×10/L vs. (9.2±4.0)×10/L, t=9.755, P<0.001], neutrophil count [(9.8±4.0)× 10/L vs.(7.1±3.9)×10/L, t=6.020, P<0.001], neutrophil percentage[(84.5±8.7)% vs.(68.2±16.0)%, t=12.754, P<0.001], C-reactive protein levels [(86.0±45.4) μg/L vs. (55.9±35.8) μg/L, t=7.614, P<0.001], serum albumin levels [(334.0±4.8) g/L vs. ( 41.0±4.3) g/L, t=16.055, P<0.001], vomiting ratio [44.6%(50/112) vs. 23.5%(47/200), χ²=14.980, P<0.001], high fever(≥39°C) ratio [16.1%(18/112) vs. 7.5%(15/200), χ²=5.577, P=0.022], the proportion of patients ≥60 years old [22.3%(25/112) vs. 13.0%(26/200), χ²=4.562, P=0.038] and previous history of appendicitis [16.1%(18/112) vs. 7.5%(15/200), χ²=5.577, P=0.022]. The above 10 variables were included in the logistic regression model for multivariate analysis. The results showed that six variables were associated with complex appendicitis. According to their strength, they were old age (≥60 years old) X1(OR=5.094), high fever (≥39°C) X2(OR=4.464), neutrophil count X6 (OR=1.269), neutrophil percentage X4 (OR=1.077), C-reactive protein level X5 (OR=1.027), and serum albumin level X3 (OR=0.763). A predictive regression model was established: P=1/[1+e], whose sensitivity and specificity of judging complex appendicitis were 76.8%(86/112) and 90.0%(180/200),respectively. Sensitivity and specificity for predictive value of complex appendicitis in further prospective validation of the model were 76.2%(48/63) and 81.1% (30/37), respectively. CONCLUSIONS Age ≥ 60 years old, body temperature ≥39°C, increased neutrophil count, neutrophil percentage and C-reactive protein levels, and hypoalbuminemia are risk factors for complex appendicitis. The establishment of predictive model may help determine complex appendicitis.
Acute Disease ; Adolescent ; Adult ; Age Factors ; Appendicitis ; diagnosis ; epidemiology ; pathology ; Case-Control Studies ; Female ; Humans ; Leukocyte Count ; Middle Aged ; Models, Statistical ; Pregnancy ; Retrospective Studies ; Risk Factors