0.05).There was significant difference in the optical density of the 43kDa protein between the patients with OAB and controls(P

BACKGROUND: Hyperbaric oxygen(HBO) is the first choice in the treatment of acute carbon monoxide(CO) poisoning. However,the mechanism of HBO in the treatment of CO poisoning,especially the mechanism in the treatmentof CO poisoning-induced delayed encephalopathy,is unclear at present.OBJECTIVE: To observe the pathological changes of hippocampal neurons in rats after acute CO poisoning to investigate the impacts of HBO therapy on Bcl-2 protein expression in hippocampal neurons in rats after CO poisoning.DESIGN: A completely randomized controlled experimental study based on the experimental animals.SETTING: Emergency department in a military medical university of Chinese PLA affiliated hospital,department of laboratory medicine in a municipal hospital,and the center of HBO therapy in a military medical university of Chinese PLA.MATERIALS: The study was conducted in the Laboratory of HBO Therapy Center,Faculty of Aerospace Medicine,the Fourth Military Medical University of Chinese PLA. Sixty male SD rats were selected.INTERVENTIONS: Sixty rats were randomly divided into 3 groups: normal control group(control group),CO poisoning group(CO group),and HBO therapy group(HBO group) with 20 rats each. Rats of each group were exposed under air or CO gas(volume fraction was 3.2 × 10-3) respectively for 60 minutes,and rats of CO-HBO group were treated by HBO. Cerebral pathological slices of hippocampus were prepared for routine HE and Bcl-2 staining to observe the characteristics of the changes of hippocampal neuronal injury and the Bcl-2protein expression on the 1st,3rd,5th and 7th day after CO poisoning.MAIN OUTCOME MEASURES: Changes of pathomorphology and Bcl-2protein expressionRESULTS: It could be seen lot of degenerated and necrotic neurons in hippocampus of rats in CO group. Degenerated and necrotic neurons decreased and the expression of Bcl-2 protein increased in CO-HBO group,especially on the 3rd and 5th day after poisoning( P ＜ 0. 05).CONCLUSION: HBO therapy can promote Bcl-2 protein expression in hippocampus after acute CO poisoning,so it can protect neurons.

Objective To summarize the diagnosis and treatment of urethral injury in children ,and to discuss the effective treat-ment method .Methods The retrospective analysis was performed on the data of 73 cases of children urethral injury in this hospital during recent 10 years .Results 66 cases were male ,7 cases were female ,aged 1-15 years old(average 7 years old) .In the male ca-ses ,there were 8 cases of anterior urethral injury ,10 cases of bulbous urethral injury ,44 cases of posterior urethral injuries and 4 cases of bladder neck injury ;in the female cases ,there were 6 cases of urethral injury and 1 case of bladder neck injury .The once cure rate of urethral realignment was 78 .6% ,which of stageⅠ urethral anastomosis was 80% and which of stage Ⅱurethral repair was 69 .6% .Conclusion Different operation modes have their advantages and disadvantages .The corresponding treatment scheme should be formulated according to the comprehensive assessment of the patient′s general condition and damage types .

Objective:To investigate the application value of computed tomography (CT) examination of lymph node short diameter in evaluating left recurrent laryngeal nerve lymph node metastasis in thoracic esophageal squamous cell carcinoma.Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 628 patients with thoracic esophageal squamous cell carcinoma who were admitted to 2 medical centers (236 cases in the Sun Yat-sen University Cancer Center and 392 cases in the Affiliated Cancer Hospital of Zhengzhou University) from October 2009 to December 2016 were collected. There were 462 males and 166 females, aged from 38 to 85 years, with a median age of 62 years. Observation indicators: (1) operation status, dissection and metastasis of left recurrent laryngeal nerve lymph node; (2) efficacy of CT examination of the left recurrent laryngeal nerve lymph node short diameter in evaluating postoperative lymph node metastasis; (3) determination of the optimal cut-off value; (4) examination results using different diagnostic criteria. Measurement data with skewed distribution were represented as M (range). Count data were described as absolute numbers or percentages. The area under curve (AUC) of receiver operating characteristic curve (ROC) was used to estimate the efficiency of detection methods. The maximum value of Youden index corresponded to the optimal cut-off point. Results:(1) Operation status, dissection and metastasis of left recurrent laryngeal nerve lymph node: among the 628 patients, there were 572 cases undergoing two-field lymph node dissection while 56 cases undergoing three-field lymph node dissection, there were 408 cases undergoing minimally invasive surgery while 220 cases undergoing open surgery. Sixty of 628 patients had left recurrent laryngeal nerve lymph node metastasis. A total of 1 666 left recurrent laryngeal nerve lymph nodes were dissected from the 628 patients, among which 75 were metastatic lymph nodes, with a metastasis rate of 4.502%(75/1 666). (2) Efficacy of CT examination of the left recurrent laryngeal nerve lymph node short diameter in evaluating postoperative lymph node metastasis: the AUC of CT examination of the left recurrent laryngeal nerve lymph node short diameter in predicting postoperative lymph node metastasis was 0.854 (95% confidence interval as 0.792 to 0.916, P<0.05). (3) Determination of the optimal cut-off value: the Youden indices were 0.556, 0.384, 0.258, 0.063 and 0.003 respectively when using 5 mm, 6 mm, 7 mm, 8 mm, 9 mm or 10 mm as the optimal cut-off value for CT examination of the left recurrent laryngeal nerve lymph node short diameter. The short diameter as 5 mm was the optimal cut-off value for CT examination of the left recurrent laryngeal nerve lymph node short diameter. (4) Examination results using different diagnostic criteria: the sensitivity, specificity, accuracy, positive predictive value, negative predictive value, cases being missed diagnosis were respectively 66.3%, 92.3%, 89.5%, 46.3%, 96.0%, 20 and 5.0%, 99.8%, 90.7%, 75.0%, 90.9%, 57 when using short diameter ≥5 mm or ≥10 mm in CT examination of the left recurrent laryngeal nerve lymph node as the diagnostic criteria for left recurrent laryngeal nerve lymph node metastasis of thoracic esophageal squamous cell carcinoma. Conclusions:CT examination of lymph node short diameter can be used to evaluate left recurrent laryngeal nerve lymph node metastasis in thoracic esophageal squamous cell carcinoma. The sensitivity, specificity and accuracy is preferable when using short diameter ≥5 mm in CT examina-tion of the left recurrent laryngeal nerve lymph node as the diagnostic criteria for left recurrent laryngeal nerve lymph node metastasis of thoracic esophageal squamous cell carcinoma.

Abstract: Objective To observe the phenotypic characteristics of 3 wild-type plague phages under different experimental environments, providing scientific evidence for the identification of phage biological characteristics and the study of their interaction with host bacteria in the future. Methods The sensitivity of 3 wild-type plague phages were detected by using liquid culture method, emisolid medium method and micro-liquid culture method based on OmniLog TM microbial identification system. Results The growth result based on LB liquid medium showed that the growth of plague phage 476 for 20-24 hours at both 28 ℃ and 37 ℃was better than that of plague phages 087 and 072204 at 37 ℃, and the growth of plague phages 087 was better than that of plague phages 072204 at 37 ℃. With the attenuated plague bacterium EV76 as the host bacterium, phage 476 was able to form visible plaque on double-layer agar medium for 20-20 hours at both 28 ℃ and 37 ℃, phages 087 and 072204 were only able to form opaque plaque on double-layer agar medium for 20-24 hours at 37 ℃. The growth results based on OmniLogTM system showed that when plague phage was lysed in EV76 strain at 33 ℃, the first row appeared as a straight line with a peak of no more than 100 in the 96-well microplate curve chart. As the phage quantity decreased, the dilution plate appeared with growth curve similar to EV76 strain in turn, and the color of tetrazolium dyes in the experimental wells gradually deepened as the phage number decreased and the host bacteria number increased. Therefore, it indicates that phage 476 was sensitively at both 28 ℃ and 37 ℃, while phage 087 and 072204 were temperature-dependent only at 37 ℃ to attenuated plague bacterium EV76. Conclusions The lysing ability of 3 wild-type plague phages are temperature-dependent, and the growth results are consistent under the three experimental conditions.

Endotoxin removal is essential for the safety of biological products. To remove endotoxin efficiently, we used polymyxin B (PMB) affinity adsorbent to remove endotoxin from protein solutions by static adsorption. We studied the effects of spacer length and ligand density of the affinity adsorbent, pH, salt type and concentration, protein type and concentration, endotoxin concentration, and additive on endotoxin removal and protein recovery. Endotoxin content and protein concentration were determined by test and Lowry assay respectively. The results showed that PMB affinity adsorbent had high capacity, high adsorption speed, high removal efficiency and good reusability. In addition, ligand density, pH, salt concentration and the isoelectric point and hydrophobicity of protein all had remarkable influence on the endotoxin removal. Under the optimal conditions, the recoveries of hemoglobin, human serum albumin and lysozyme were 87.2%, 73.4% and 97.3%, respectively, and the corresponding endotoxin removal rates 99.8%, 97.9% and 99.7%, respectively. This study illustrated the effects of solution conditions on the efficiency of endotoxin removal and protein recovery, and would provide useful reference for the efficient removal of endotoxin from biological products.
Adsorption ; Chromatography, Affinity ; methods ; Drug Contamination ; prevention & control ; Endotoxins ; isolation & purification ; Hemoglobins ; isolation & purification ; Polymyxin B ; chemistry ; Proteins ; isolation & purification ; Pyrogens ; isolation & purification ; Serum Albumin ; isolation & purification ; Solutions

Objective:To find out predictors of the testicular ischemia caused by incarcerated inguinal hernia and evaluate the ischemic injury of the testis more accurately, which can indicate testicle exploration in time or prevent unnecessary testicle exploration.Methods:Pediatric patients (median: 9 months) undergoing operation of unilateral incarcerated inguinal hernia and ipsilateral testicular exploration from 1 Jul. 2013 to 30 Jun. 2019 were retrospectively investigated. Age at surgery, incarcerate duration, degree of intestinal and testicular injury, times of manual reduction and preoperative ultrasound data were collected. Statistical analysis was performed by SAS 9.4 (Copyright ? 2016 SAS Institute Inc.Cary, NC, USA) .Results:460 patients (median: 9 months) , of which 57 (12.39%) (median: 1.4 months, interquartile range 0.8-10.7 months) had severe testicular injury, and their average incarceration time was (23.9±9.3) h. Univariate logistic regression revealed that increased times of manual reduction, ultrasound scores, incarcerate duration and degree of intestinal injury were positively correlated with the degree of testicular ischemia, while age at surgery was negatively correlated with the degree of testicular ischemia ( P<0.05) . A model for calculating the probability of severe testicular ischemia injury was established: P= through multivariate analysis with backward stepwise logistic regression and 10-fold cross-validation was used for preliminary verification of the model. Conclusion:This study provides a relative reliable model to predict the risk of irreversible testicular ischemia due to incarcerated inguinal hernia using readily available clinical characteristics in young pediatrics with testicular ischemia.

Objective To analyze the frequency of pelvis peristalsis in children with hydronephrosis and its correlation with postoperative percentage of improvement in anteroposterior diameter (PI－APD).Methods From Oc-tober 2015 to September 2017,the data of the children with unilateral ureteropelvic junction obstruction (UPJO)hydro-nephrosis who underwent laparoscopic pyeloplasty were collected,and their pelvis peristalsis frequencies of 2 min were recorded after exposing the pelvis,while the APD and PI－APD were followed 1 year posto-peratively.The degrees of hydronephrosis were divided into mild,moderate and severe according to the Society of Fetal Urology(SFU)classifica-tion system.ANOVA was used to compare the pelvis peristalsis frequency and postoperative PI－APD between different degrees of hydronephrosis.Spearman analysis was used to analyze the correlation between pelvis peristalsis frequency and degree of hydronephrosis.Meanwhile,Pearson test was used to analyze the correlation between pelvis peristalsis fre-quency,hydronephrosis degree and PI－APD.Results A total of 50 children were included,of which 36 patients got follow－up for 1 year postoperatively.Pelvis 2 min peristalsis frequency between different degrees of hydronephrosis [mild:(8.1 ± 3. 3)times;moderate:(6.3 ± 3.5)times;severe:(7.8 ± 3.9)times]had no significant difference (F＝0.65,P＝0. 527);no statistical correlation was observed between pelvis peristalsis frequency and degree of hydro-nephrosis or PI－APD (all P ＞0.05).Preoperative hydronephrosis degree was positively correlated with PI －APD (r＝0.54,P＝0. 001).PI－APD in severe hydronephrosis (0.48 ± 0.29)was significantly higher than that of mild hydronephrosis (0.21 ± 0.20)(P ＝0.001 ). Conclusions For children with different degrees of hydronephrosis caused by UPJO,there was no significant difference in the pelvis peristalsis frequency recorded during laparoscopic pyeloplasty.Pelvis peristalsis frequency recorded during surgery was not correlated with PI －APD,but preoperative hydronephrosis degree was positively correlated with postoperative PI－APD.

Objective To observe the value of micro flow imaging(MFI)for diagnosing closed liver trauma in pigs.Methods A self-made impactor was used to impact the liver of 15 healthy pigs under general anesthesia and establish pig models of closed liver trauma.Conventional ultrasound,MFI and contrast-enhanced ultrasound(CEUS)were used to observe the location,extent and degree of liver trauma.Afterwards,the pigs were executed and the livers were removed,and the liver lesions were observed.According gross pathological findings,the diagnostic efficacy of MFI was analyzed.Results Gross pathology showed a total of 29 liver traumas in 15 pigs.The accuracy rate of conventional ultrasound,MFI and CEUS for diagnosing liver trauma in pigs was 37.93％(11/29),82.76％(24/29)and 96.55％(28/29),respectively.Significant difference was found between conventional ultrasound and MFI,also between conventional ultrasound and CEUS(both P＜0.05),but not between MFI and CEUS(P＞0.05).MFI showed no obvious blood flow signal in more than half pig liver trauma sites,while showed"spring like"in 1 pig and scattered blood flow signals in 5 pigs.On CEUS,liver trauma sites in pigs present as low or no perfusion,while"jet like"contrast agent overflow from the liver was noticed in 1 pig and"spring like"contrast agent overflow in 5 pigs,forming liquid dark areas around the liver or in abdominal cavity.Conclusion MFI had certain value for diagnosing closed liver trauma in pigs,hence being comparable to CEUS.

OBJECTIVETo observe the histological features of tumor-bearing tissues formed by human fibroblasts after reprograming by spermatogonial stem cell self-renewal key regulating gene Piwil2 (Piwil2-iCSC).

METHODSPiwil2-iCSC tumor spheroids-like colonies were selected for tumor formation assay in four nude mice. Pathological features of Piwil2-iCSC tumors were observed by histology. Stem cell markers and common triploblastic markers were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) assay and immunohistochemistry. Germ cell tumor markers were detected by immunohistochemical examination.

RESULTSTwo weeks after inoculation, subcutaneous tumors were formed in all the four nude mice with a tumor formation rate of 100%. In the Piwil2-iCSC tumor tissues, Piwil2-GFP(+) cells showed high-density nuclear expression and were widely observed in DAPI-stained sections. Numerous mitotic figure of the neoplastic cells were seen (>10 cells/field of vision under high magnification) in HE-stained sections. Enlarged abnormal cell nuclei were observed. RT-PCR assay showed that Piwil2-iCSC tumors still expressed Piwil2 and some self-renewal and pluripotent markers of stem cells and some markers of triploblastic differentiation. Immunohistochemical staining showed that the tumors expressed stem cell markers, triploblastic markers and germ cell tumor markers AFP and HCG.

CONCLUSIONSPiwil2-iCSC tumors are probably undifferentiated embryonic small cell carcinoma, most likely to be immature teratoma, mixed with yolk sac tumor and choriocarcinoma components. It can be used as a useful model for the research of origin or genesis mechanism of cancer stem cells and the treatment of relevant tumors.

Adult Stem Cells ; Animals ; Argonaute Proteins ; genetics ; Cellular Reprogramming Techniques ; Choriocarcinoma ; pathology ; Endodermal Sinus Tumor ; pathology ; Fibroblasts ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Mice ; Mice, Nude ; Neoplasms, Germ Cell and Embryonal ; chemistry ; genetics ; pathology ; Neoplastic Stem Cells ; chemistry ; pathology ; Real-Time Polymerase Chain Reaction ; Spheroids, Cellular ; Teratoma ; pathology ; Time Factors